Alpha-2 agonist polymeric drug delivery systems

ABSTRACT

Biocompatible intraocular implants include an alpha-2 adrenergic receptor agonist and a polymer associated with the alpha-2 adrenergic receptor agonist to facilitate release of the alpha-2 adrenergic receptor agonist into an eye for an extended period of time. The alpha-2 adrenergic receptor agonist may be associated with a biodegradable polymer matrix, such as a matrix of a two biodegradable polymers. The implants can be placed in an eye to treat one or more ocular conditions, such as an ocular vasculopathy or glaucoma, including reduction of an elevated intraocular pressure.

CROSS REFERENCE

This application is a continuation of copending application Ser. No.11/394,765, filed Mar. 31, 2006, which is a continuation in part ofapplication Ser. No. 11/119,021, filed Apr. 29, 2005, now issued as U.S.Pat. No. 8,293,741, which is continuation in part of application Ser.No. 10/836,911 filed Apr. 30, 2004, now abandoned. The entire contentsof these applications are incorporated herein by reference.

BACKGROUND

The present invention generally relates to devices and methods to treatan eye of a patient, and more specifically to intraocular implants thatprovide extended release of a therapeutic agent to an eye in which theimplant is placed, and to methods of making and using such implants, forexample, to treat ocular vasculopathies, or to generally improve vision.

Brimonidine, 5-bromo-6-(2-imidazolidinylideneamino) quinoxaline, is analpha-2-selective adrenergic receptor agonist that is effective in thetreatment of open-angle glaucoma by decreasing aqueous humor productionand increasing uveoscleral outflow. Brimonidine is available in twochemical forms, brimonidine tartrate and brimonidine free base.Brimonidine tartrate (Alphagan P®) is commercially available fromAllergan for treating glaucoma. Topical ocular brimonidine formulation,0.15% Alphagan P® (Allergan, Irvine, Calif.), is currently commerciallyavailable for treatment of open-angle glaucoma. The solubility ofbrimonidine tartrate in water is 34 mg/mL, while the solubility ofbrimonidine freebase is negligible in water.

Recent studies have suggested that brimonidine can promote survival ofinjured retinal ganglion nerve cells by activation of thealpha-2-adrenoceptor in the retina and/or optic nerve. For example,brimonidine can protect injured neurons from further damage in severalmodels of ischemia and glaucoma. See e.g. U.S. Pat. Nos. 5,856,329;6,194,415; 6,248,741, and; 6,465,464.

Glaucoma-induced ganglion cell degeneration is one of the leading causesof blindness. This indicates that brimonidine can be utilized in a newtherapeutic approach to glaucoma management in which neuroprotection andintraocular pressure reduction are valued outcomes of the therapeuticregimen. For brimonidine to protect the optic nerve, however, it musthave access to the posterior segment of the eye at therapeutic levels.Currently available techniques for administering brimonidine to theposterior chamber of the eye are not sufficient to address this issue.It has been reported that intravitreal injection of brimonidine may havea neuroprotective effect. Gao H., et al., Up-regulation of brain-derivedneurotrophic factor expression by brimonidine in rat retinal ganglioncells, Arch Opthal 2002 June; 120(6): 797-803.

Biocompatible implants for placement in the eye have been disclosed in anumber of patents, such as U.S. Pat. Nos. 4,521,210; 4,853,224;4,997,652; 5,164,188; 5,443,505; 5,501,856; 5,766,242; 5,824,072;5,869,079; 6,074,661; 6,331,313; 6,369,116; and 6,699,493.

It would be advantageous to provide eye implantable drug deliverysystems, such as intraocular implants, and methods of using suchsystems, that are capable of releasing a therapeutic agent at asustained or controlled rate for extended periods of time and in amountswith few or no negative side effects.

SUMMARY

The present invention provides new drug delivery systems, and methods ofmaking and using such systems, for extended or sustained drug releaseinto an eye, for example, to achieve one or more desired therapeuticeffects. The drug delivery systems are in the form of implants orimplant elements that may be placed in an eye. The present systems andmethods advantageously provide for extended release times of one or moretherapeutic agents. Thus, the patient in whose eye the implant has beenplaced receives a therapeutic amount of an agent for a long or extendedtime period without requiring additional administrations of the agent.For example, the patient has a substantially consistent level oftherapeutically active agent available for consistent treatment of theeye over a relatively long period of time, for example, on the order ofat least about one week, such as between about two and about six monthsafter receiving an implant. Such extended release times facilitateobtaining successful treatment results.

Intraocular implants in accordance with the disclosure herein comprise atherapeutic component and a drug release sustaining component associatedwith the therapeutic component. In accordance with the presentinvention, the therapeutic component comprises, consists essentially of,or consists of, an alpha-2 adrenergic receptor agonist. The alpha-2adrenergic receptor agonist may be an agonist or agent that selectivelyactivates alpha-2 adrenergic receptors, for example by binding to analpha-2 adrenergic receptor, relative to other types of adrenergicreceptors, such as alpha-1 adrenergic receptors. The selectiveactivation can be achieved under different conditions, but preferably,the selective activation is determined under physiological conditions,such as conditions associated with an eye of a human or animal patient.The drug release sustaining component is associated with the therapeuticcomponent to sustain release of an amount of the alpha-2 adrenergicreceptor agonist into an eye in which the implant is placed. The amountof the alpha-2 adrenergic receptor agonist is released into the eye fora period of time greater than about one week after the implant is placedin the eye and is effective in preventing or reducing ocularvasculopathies, such as vascular occlusions.

In one embodiment, the intraocular implants comprise an alpha-2adrenergic receptor agonist and a biodegradable polymer matrix. Thealpha-2 adrenergic receptor agonist is associated with a biodegradablepolymer matrix that degrades at a rate effective to sustain release ofan amount of the agonist from the implant for a time sufficient toreduce or prevent an ocular vascular occlusion. The intraocular implantis biodegradable or bioerodible and provides a sustained release of thealpha-2 adrenergic receptor agonist in an eye for extended periods oftime, such as for more than one week, for example for about three monthsor more and up to about six months or more. In certain implants, thealpha-2 adrenergic receptor agonist is released for about 30-35 days orless. In other implants, the alpha-2 adrenergic receptor agonist isreleased for 40 days or more.

The biodegradable polymer component of the foregoing implants may be amixture of biodegradable polymers, wherein at least one of thebiodegradable polymers is a polylactic acid polymer having a molecularweight less than 64 kiloDaltons (kD). Additionally or alternatively, theforegoing implants may comprise a first biodegradable polymer of apolylactic acid, and a different second biodegradable polymer of apolylactic acid. Furthermore, the foregoing implants may comprise amixture of different biodegradable polymers, each biodegradable polymerhaving an inherent viscosity in a range of about 0.3 deciliters/gram(dl/g) to about 1.0 dl/g.

The alpha-2 adrenergic receptor agonist of the implants disclosed hereinmay include quinoxaline derivatives, or other agonists that areeffective in treating ocular conditions. One example of a suitablequinoxaline derivative is brimonidine or brimonidine tartrate. Inaddition, the therapeutic component of the present implants may includeone or more additional and different therapeutic agents that may beeffective in treating an ocular condition.

A method of making the present implants involves combining or mixing thealpha-2 adrenergic receptor agonist with a biodegradable polymer orpolymers. The mixture may then be extruded or compressed to form asingle composition. The single composition may then be processed to formindividual implants suitable for placement in an eye of a patient.

The implants may be placed in an ocular region to treat a variety ofocular conditions, including conditions such as ocular vasculopathiesthat affect an anterior region or posterior region of an eye. Forexample, the implants may be used to treat many conditions of the eye,including, without limitation, conditions associated with vascularocclusion.

Kits in accordance with the present invention may comprise one or moreof the present implants, and instructions for using the implants. Forexample, the instructions may explain how to administer the implants toa patient, and types of conditions that may be treated with theimplants.

The present invention also encompasses a biodegradable intraocularimplant for improving vision. The implant can comprise an alpha-2adrenergic receptor agonist and a biodegradable polymer. The implantreleases the alpha-2 adrenergic receptor agonist from the polymer, uponintravitreal placement of the implant, in an amount effective to improvethe vision of the eye in which the implant is placed. The alpha-2adrenergic receptor agonist can be a quinoxaline, such as a(2-imidazolin-2-ylamino) quinoxaline, a5-bromo-6-(2-imidazolin-2-ylamino) quinoxaline, and derivatives thereofand mixtures thereof. Thus, the alpha-2 adrenergic receptor agonist canbe a brimonidine or salts thereof or mixtures thereof. For example, thealpha-2 adrenergic receptor agonist can be brimonidine tartrate.

The alpha-2 adrenergic receptor agonist can be dispersed within thebiodegradable polymer of the implant. The biodegradable polymer cancomprise a mixture of a first biodegradable polymer of polylactic acid,and a different second biodegradable polymer of polylactic acid. Thepolymer can release drug at a rate effective to sustain release of anamount of the alpha-2 adrenergic receptor agonist from the implant formore than one month or for more that forty days or for less than thirtyfive days from the time the implant is placed in the vitreous of theeye.

An embodiment of the present invention is a method of making abiodegradable intraocular implant by extruding a mixture of an alpha-2adrenergic receptor agonist and a biodegradable polymer component toform a biodegradable material that releases drug at a rate effective tosustain release of an amount of the alpha-2 adrenergic receptor agonistfrom the implant for a time effective to improve vision in an eye inwhich the implant is placed.

A further embodiment of the present invention is a method for improvingor for maintaining vision by placing in the vitreous of an eye abiodegradable intraocular implant comprising an alpha-2 adrenergicreceptor agonist associated with a biodegradable polymer, therebyimproving or maintaining vision. This method can be used to treat anocular condition such as: macular degeneration, macular edema, retinalarterial occlusive disease, central retinal vein occlusion, disseminatedintravascular coagulopathy, branch retinal vein occlusion, hypertensivefundus changes, ocular ischemic syndrome, retinal arterialmicroaneurysms, hemi-retinal vein occlusion, central retinal arteryocclusion, branch retinal artery occlusion, carotid artery disease(cad), eales disease, vasculopathies associated with diabetes,Non-Exudative Age Related Macular Degeneration, Exudative Age RelatedMacular Degeneration, Choroidal Neovascularization, DiabeticRetinopathy, Acute Macular Neuroretinopathy, Central SerousChorioretinopathy, Cystoid Macular Edema, Diabetic Macular Edema, AcuteMultifocal Placoid Pigment Epitheliopathy, Behcet's Disease, BirdshotRetinochoroidopathy, Syphilis, Lyme, Tuberculosis, Toxoplasmosis,Intermediate Uveitis, Multifocal Choroiditis, Multiple Evanescent WhiteDot Syndrome, Ocular Sarcoidosis, Posterior Scleritis, SerpiginousChoroiditis, Subretinal Fibrosis and Uveitis Syndrome,Vogt-Koyanagi-Harada Syndrome, Coat's Disease, ParafovealTelangiectasia, Papillophlebitis, Frosted Branch Angiitis, Sickle CellRetinopathy and other Hemoglobinopathies, Angioid Streaks, FamilialExudative Vitreoretinopathy, Sympathetic Ophthalmia, Uveitic RetinalDisease, Retinal Detachment, Trauma, Laser, photodynamic therapy,Photocoagulation, Hypoperfusion During Surgery, Radiation Retinopathy,Bone Marrow Transplant Retinopathy, Proliferative Vitreal Retinopathyand Epiretinal Membranes, Proliferative Diabetic Retinopathy, OcularHistoplasmosis, Ocular Toxocariasis, Presumed Ocular HistoplasmosisSyndrome, Endophthalmitis, Toxoplasmosis, Retinal Diseases Associatedwith HIV Infection, Choroidal Disease Associated with HIV Infection,Uveitic Disease Associated with HIV Infection, Viral Retinitis, AcuteRetinal Necrosis, Progressive Outer Retinal Necrosis, Fungal RetinalDiseases, Ocular Syphilis, Ocular Tuberculosis, Diffuse UnilateralSubacute Neuroretinitis, Myiasis, Retinitis Pigmentosa, SystemicDisorders with Associated Retinal Dystrophies, Congenital StationaryNight Blindness, Cone Dystrophies, Stargardt's Disease and FundusFlavimaculatus, Best's Disease, Pattern Dystrophy of the RetinalPigmented Epithelium, X-Linked Retinoschisis, Sorsby's Fundus Dystrophy,Benign Concentric Maculopathy, Bietti's Crystalline Dystrophy,pseudoxanthoma elasticum, Retinal Detachment, Macular Hole, GiantRetinal Tear, Retinal Disease Associated with Tumors, CongenitalHypertrophy of the RPE, Posterior Uveal Melanoma, Choroidal Hemangioma,Choroidal Osteoma, Choroidal Metastasis, Combined Hamartoma of theRetina and Retinal Pigmented Epithelium, Retinoblastoma,Vasoproliferative Tumors of the Ocular Fundus, Retinal Astrocytoma,Intraocular Lymphoid Tumors, Punctate Inner Choroidopathy, AcutePosterior Multifocal Placoid Pigment Epitheliopathy, Myopic RetinalDegeneration, and Acute Retinal Pigment Epithelitis.

Notably, the method can improve vision in a normal eye. A normal eye isan eye which is not diseased or damaged. For example, the method canimprove vision (as by improving visual acuity) in a normal eye by up toabout 56%. The method can also improve vision in an eye with an ocularcondition. For example, the method can improve vision in an eye with anocular condition by up to about 23%. The ocular condition can be avasculopathy. Alternately, the ocular condition can be due to anelevated intraocular pressure and/or the ocular condition can be aretinal ischemic injury.

The implant can release the alpha-2 adrenergic receptor agonist from thepolymer, upon intravitreal placement of the implant, for a period ofabout ninety days. Significantly, the alpha-2 adrenergic receptoragonist can be retained in the retina for a period of time longer thanit is retained in the vitreous. An embodiment of the present inventionis a method for improving, maintaining, restoring or repairing vision,the method comprising the step of placing in the vitreous of an eye abiodegradable intraocular implant comprising a brimonidine associatedwith a biodegradable polymer, thereby improving, maintaining, restoringor repairing vision.

An embodiment of our invention is a biodegradable intraocular implantcomprising an alpha-2 adrenergic receptor agonist and a biodegradablepolymer, wherein the biodegradable polymer comprises an ester end-cappedbiodegradable polymer and an acid end-capped biodegradable polymer. Theimplant can comprise from about 10% to about 91% ester end-cappedbiodegradable polymer, from about 5 wt % to about 40 wt % acidend-capped biodegradable polymer, and from about 4 wt % to about 50 wt %alpha-2 adrenergic receptor agonist. Preferably, the implant cancomprise from about 45% to about 80% ester end-capped biodegradablepolymer, from about 10 wt % to about 40 wt % acid end-cappedbiodegradable polymer, and about 10 wt % to about 15 wt % alpha-2adrenergic receptor agonist. More preferably, the implant can compriseabout 88 wt % ester end-capped biodegradable polymer, about 10 wt % acidend-capped biodegradable polymer, and about 12 wt % alpha-2 adrenergicreceptor agonist. Most preferably, the implant can comprise from about53 wt % to about 73% ester end-capped biodegradable polymer, from about15 wt % to about 35 wt % acid end-capped biodegradable polymer, and fromabout 9 wt % to about 12 wt % alpha-2 adrenergic receptor agonist.

The biodegradable polymer of the implant can comprise more than oneester end-capped biodegradable polymer. Alternately, the biodegradablepolymer of the implant can comprise more than one acid end-cappedbiodegradable polymer. The implant can have no or a nominal lag timeafter ocular implantation or insertion of the implant before release ofa therapeutically effective amount of the alpha-2 adrenergic receptoragonist from the implant occurs. The implant comprise greater than orequal to 4 weight percent (wt %) of a biologically active alpha-2adrenergic receptor agonist and the implant preferably does not includeany pore forming additives, release rate modulators or release ratemodifiers. The implant can exhibit a sustained release of the alpha-2adrenergic receptor agonist from the biodegradable polymeric matrix overa period of at least 115 days. Additionally, the implant can exhibit asubstantially linear release of the alpha-2 adrenergic receptor agonistfrom the biodegradable polymeric matrix of the implant over a period oftime of from about 20 days to about 50 days.

A preferred embodiment of a biodegradable intraocular implant within thescope of our invention can comprise an alpha-2 adrenergic receptoragonist, and a biodegradable polymer, wherein the biodegradable polymercomprises an ester end-capped biodegradable polymer and an acidend-capped biodegradable polymer, wherein the implant comprises fromabout 40% to about 91% of at least two different ester end-cappedbiodegradable polymers, from about 5 wt % to about 40 wt % acidend-capped biodegradable polymer, and from about 4 wt % to about 20 wt %alpha-2 adrenergic receptor agonist.

Our invention also includes a process for making a biodegradableintraocular implant by mixing an alpha-2 adrenergic receptor agonist anda biodegradable polymer, wherein the biodegradable polymer comprises anester end-capped biodegradable polymer and an acid end-cappedbiodegradable polymer; heating the mixture, and; extruding the heatedmixture, to thereby make a biodegradable intraocular implant.

An implant within the scope of our invention can be an extruded filamentwith a diameter of about 0.5 mm, a length of about 6 mm and a weight ofabout 1 mg. The alpha-2 adrenergic receptor agonist can be homogenouslydistributed throughout the implant.

Our implants can be used to treat ocular conditions by intraocularadministration of a biodegradable intraocular implant comprising analpha-2 adrenergic receptor agonist and a biodegradable polymer, whereinthe biodegradable polymer comprises an ester end-capped biodegradablepolymer and an acid end-capped biodegradable polymer. The alpha-2adrenergic receptor agonist can be selected from the group consisting ofbrimonidine, salts thereof, and mixtures thereof.

In another embodiment of our invention a biodegradable intraocularimplant can comprise a plurality of forms of an alpha-2 adrenergicreceptor agonist and a biodegradable polymer. The alpha-2 adrenergicreceptor agonist can be a brimonidine and the brimonidine can be presentin two forms in the implant. The two forms of brimonidine present in theimplant can be brimonidine free base and brimonidine tartrate. Such andimplant can comprises from about 50 wt % to about 70% ester end-cappedbiodegradable polymer, from about 1 wt % to about 49 wt % brimonidinefree base and from about 1 wt % to about 49 wt % brimonidine tartrate.Alternately, the implant can comprises from about 50 wt % to about 60%ester end-capped biodegradable polymer, from about 1 wt % to about 49 wt% brimonidine free base and from about 1 wt % to about 49 wt %brimonidine tartrate. More preferably, the implant can comprise fromabout 50 wt % to about 70% ester end-capped biodegradable polymer, fromabout 10 wt % to about 30 wt % brimonidine free base and from about 10wt % to about 30 wt % brimonidine tartrate. In most preferred embodimentthe implant can comprise from about 55 wt % to about 65% esterend-capped biodegradable polymer, from about 15 wt % to about 20 wt %brimonidine free base and from about 15 wt % to about 20 wt %brimonidine tartrate, for example the implant can comprise about 65 wt %ester end-capped biodegradable polymer, about 18 wt % brimonidine freebase and about 18 wt % brimonidine tartrate. The implant of claim 21,wherein the biodegradable polymer comprises more than one esterend-capped biodegradable polymer. And the implant can have no bursteffect and no or a nominal lag time after ocular implantation orinsertion of the implant before release of a therapeutically effectiveamount of the alpha-2 adrenergic receptor agonist from the implantoccurs. Additionally, the implant can exhibit a sustained release of thealpha-2 adrenergic receptor agonist from the biodegradable polymericmatrix over a period of at least 60 days. Furthermore, the implant canexhibits a substantially linear release of the alpha-2 adrenergicreceptor agonist from the biodegradable polymeric matrix of the implantover a period of time of from about 20 days to about 50 days.

A preferred embodiment of our invention can comprise a brimonidine freebase; a brimonidine tartrate, and an ester end-capped biodegradablepolymer, wherein the implant comprises from about 50 wt % to about 70%of the ester end-capped biodegradable polymer, from about 1 wt % toabout 49 wt % of the brimonidine free base and from about 1 wt % toabout 49 wt % of the brimonidine tartrate.

Our invention encompasses a process for making a biodegradableintraocular implant comprising (a) mixing a plurality of forms ofalpha-2 adrenergic receptor agonist and a biodegradable polymer; (b)heating the mixture, and; (c) extruding the heated mixture, to therebymake a biodegradable intraocular implant. The implant can be extruded asa filament with a diameter of about 0.5 mm, a length of about 6 mm and aweight of about 1 mg. The implant can also be made by a directcompression or solvent extraction method. The shape of the implant canalso be as a tablet, pellet or rod.

Finally, our invention encompasses a method of treating a symptom ofglaucoma by placing a biodegradable intraocular implant comprising analpha-2 adrenergic receptor agonist associated with a biodegradablepolymer into the vitreous of an eye, thereby treating a symptom of theglaucoma. The symptom of the glaucoma can be reduced for at least about35 days after intravitreal placement of the implant. The symptom of theglaucoma treated can be an elevated intraocular pressure.

Additional aspects and advantages of the present invention are set forthin the following description and claims, particularly when considered inconjunction with the accompanying drawings.

DRAWINGS

FIG. 1 is a graph showing the cumulative release profiles forbiodegradable brimonidine tartrate containing implants as determined in0.9% phosphate buffered saline at 37 degrees Celsius.

FIG. 2 is a graph similar to FIG. 1 showing the cumulative releaseprofiles for biodegradable brimonidine free base containing implantswith different combinations of biodegradable polymers.

FIG. 3 is a graph similar to FIG. 1 showing the cumulative releaseprofiles for biodegradable brimonidine tartrate containing implantshaving different concentrations of brimonidine tartrate.

FIG. 4 is a graph similar to FIG. 3 showing the cumulative releaseprofiles for biodegradable brimonidine tartrate containing implantshaving different concentrations of brimonidine tartrate and polymericblends.

FIG. 5 is a graph similar to FIG. 4 showing the cumulative releaseprofiles for biodegradable brimonidine free base containing implantshaving different concentrations of brimonidine tartrate and polymericblends.

FIG. 6 is a graph showing the cumulative release profiles forbrimonidine tartrate containing implants (wafers) having differentconcentrations of brimonidine tartrate and polymeric combinations.

FIG. 7 is a graph similar to FIG. 6 showing the cumulative releaseprofiles for biodegradable brimonidine free base containing implantshaving a different concentration of brimonidine tartrate and polymericblends.

FIG. 8 is a graph similar to FIG. 4 showing the cumulative releaseprofiles for biodegradable brimonidine free base containing implantshaving a different concentration of brimonidine tartrate and polymericblends.

FIG. 9 is a graph similar to FIG. 5 showing the cumulative releaseprofiles for biodegradable brimonidine free base containing waferimplants.

FIG. 10 is a graph showing the delay in filling of sodium fluoresceinduring angiography following branch retinal vein occlusion (BRVO) versustime in monkeys that have received brimonidine tartrate containingbiodegradable implants or placebo implants.

FIG. 11 is a graph of foveal thickness as a function of time in monkeysthat have received brimonidine tartrate containing biodegradableimplants or placebo implants and experienced BRVO.

FIG. 12 is a graph of intraocular pressure as a function of time inmonkeys that have received brimonidine tartrate containing biodegradableimplants or placebo implants and experienced BRVO.

FIG. 13 is a graph of the superior/inferior percent response to amultifocal ERG as a function of time in monkeys that have receivedbrimonidine tartrate containing biodegradable implants or placeboimplants and experienced BRVO.

FIG. 14 is a graph of blood flow as a function of time in monkeys thathave received brimonidine tartrate containing biodegradable implants orplacebo implants and experienced BRVO.

FIG. 15 is a bar graph showing the effect upon visual acuity (as apercent of the visual acuity of the untreated [control] left eye)(Y-axis) in normal rabbit right eyes two weeks after intravitrealadministration of either a brimonidine implant or a placebo implant(X-axis).

FIG. 16 is a bar graph showing the effect upon visual acuity (as apercent of the visual acuity of the untreated [control] left eye)(Y-axis) three weeks after intravitreal administration of either abrimonidine implant or of a placebo implant in the right eyes and oneweek after VEGF induced injury of the same right eyes (X-axis).

FIG. 17 is a bar graph showing the effect upon visual acuity (as apercent of the visual acuity of the untreated [control] right eye)(Y-axis) twelve weeks after intravitreal administration of either abrimonidine implant or of a placebo implant in the right eyes and elevenmonths after induced ischemic injury of the same right eyes X-axis).

FIG. 18 is a graph which illustrates three different possible releaseprofiles for the release of an active agent from an implant. Amount ofan active agent released is shown on the Y-axis while the X-axisrepresents time after intraocular placement of the implant.

FIG. 19 is a graph showing the percent of the total amount ofbrimonidine released (Y-axis) versus time (X-axis) in days for a periodof 21 days in vitro for the three Table 4 implants.

FIG. 20 is a graph showing the percent of the total amount ofbrimonidine released (Y-axis) versus time in days (X-axis) for a periodof 151 days in vitro for the Table 4 implant 7746-073.

FIG. 21 is a graph showing the percent of the total amount ofbrimonidine released (Y-axis) versus time (X-axis) in days for a periodof 14-26 days in vitro for the seven the Table 5 implants.

FIG. 22 is a graph showing the percent of the total amount ofbrimonidine released (Y-axis) versus time (X-axis) in days over a periodof 60 days in vitro for the six Table 6 implants.

FIG. 23 is a graph showing intraocular pressure (in mm Hg) on the Y-axisand time in weeks on the X-axis after intravitreal placement of theExample 10 brimonidine implant in hypertensive rabbit eyes.

DESCRIPTION

As described herein, controlled and sustained administration of atherapeutic agent through the use of one or more intraocular implantsmay improve treatment of undesirable ocular conditions. The implantscomprise a pharmaceutically acceptable polymeric composition and areformulated to release one or more pharmaceutically active agents, suchas alpha-2 adrenergic receptor agonists, over an extended period oftime. The implants are effective to provide a therapeutically effectivedosage of the agent or agents directly to a region of the eye to treator prevent one or more undesirable ocular conditions. Thus, with asingle administration, therapeutic agents will be made available at thesite where they are needed and will be maintained for an extended periodof time, rather than subjecting the patient to repeated injections or,in the case of self-administered drops, ineffective treatment with onlylimited bursts of exposure to the active agent or agents.

An intraocular implant in accordance with the disclosure hereincomprises a therapeutic component and a drug release sustainingcomponent associated with the therapeutic component. In accordance withthe present invention, the therapeutic component comprises, consistsessentially of, or consists of, an alpha-2 adrenergic receptor agonist.The drug release sustaining component is associated with the therapeuticcomponent to sustain release of a therapeutically effective amount ofthe alpha-2 adrenergic receptor agonist into an eye in which the implantis placed. The therapeutic amount of the alpha-2 adrenergic receptoragonist is released into the eye for a period of time greater than aboutone week after the implant is placed in the eye.

DEFINITIONS

For the purposes of this description, we use the following terms asdefined in this section, unless the context of the word indicates adifferent meaning.

“About” means plus or minus ten percent of the value so qualified.

“Biocompatible” means that there is an insignificant inflammatoryresponse upon contact of the biocompatible material with an oculartissue.

“Effective amount” as applied to an active agent means that amount ofthe compound which is generally sufficient to effect a desired change inthe subject.

“Intraocular implant” means a device or element that is structured,sized, or otherwise configured to be placed in an eye. Intraocularimplants are generally biocompatible with physiological conditions of aneye and do not cause adverse side effects. Intraocular implants may beplaced in an eye without disrupting vision of the eye.

“Therapeutic component” means a portion of an intraocular implantcomprising one or more therapeutic agents or substances used to treat amedical condition of the eye. The therapeutic component may be adiscrete region of an intraocular implant, or it may be homogenouslydistributed throughout the implant. The therapeutic agents of thetherapeutic component are typically ophthalmically acceptable, and areprovided in a form that does not cause adverse reactions when theimplant is placed in an eye.

“Drug release sustaining component” means a portion of the intraocularimplant that is effective to provide a sustained release of thetherapeutic agents of the implant. A drug release sustaining componentmay be a biodegradable polymer matrix, or it may be a coating covering acore region of the implant that comprises a therapeutic component.

“Associated with” means mixed with, dispersed within, coupled to,covering, or surrounding.

“Ocular region” or “ocular site” means any area of the eyeball,including the anterior and posterior segment of the eye, and whichgenerally includes, but is not limited to, any functional (e.g., forvision) or structural tissues found in the eyeball, or tissues orcellular layers that partly or completely line the interior or exteriorof the eyeball. Specific examples of areas of the eyeball in an ocularregion include the anterior chamber, the posterior chamber, the vitreouscavity, the choroid, the suprachoroidal space, the conjunctiva, thesubconjunctival space, the episcleral space, the intracorneal space, theepicorneal space, the sclera, the pars plana, surgically-inducedavascular regions, the macula, and the retina.

“Ocular condition” means a disease, ailment or condition which affectsor involves the eye or one of the parts or regions of the eye. Broadlyspeaking the eye includes the eyeball and the tissues and fluids whichconstitute the eyeball, the periocular muscles (such as the oblique andrectus muscles) and the portion of the optic nerve which is within oradjacent to the eyeball.

An anterior ocular condition is a disease, ailment or condition whichaffects or which involves an anterior (i.e. front of the eye) ocularregion or site, such as a periocular muscle, an eye lid or an eye balltissue or fluid which is located anterior to the posterior wall of thelens capsule or ciliary muscles. Thus, an anterior ocular conditionprimarily affects or involves the conjunctiva, the cornea, the anteriorchamber, the iris, the posterior chamber (behind the retina but in frontof the posterior wall of the lens capsule), the lens or the lens capsuleand blood vessels and nerve which vascularize or innervate an anteriorocular region or site.

Thus, an anterior ocular condition can include a disease, ailment orcondition, such as for example, aphakia; pseudophakia; astigmatism;blepharospasm; cataract; conjunctival diseases; conjunctivitis; cornealdiseases; corneal ulcer; dry eye syndromes; eyelid diseases; lacrimalapparatus diseases; lacrimal duct obstruction; myopia; presbyopia; pupildisorders; refractive disorders and strabismus. Glaucoma can also beconsidered to be an anterior ocular condition because a clinical goal ofglaucoma treatment can be to reduce a hypertension of aqueous fluid inthe anterior chamber of the eye (i.e. reduce intraocular pressure).

A posterior ocular condition is a disease, ailment or condition whichprimarily affects or involves a posterior ocular region or site such aschoroid or sclera (in a position posterior to a plane through theposterior wall of the lens capsule), vitreous, vitreous chamber, retina,optic nerve (i.e. the optic disc), and blood vessels and nerves whichvascularize or innervate a posterior ocular region or site.

Thus, a posterior ocular condition can include a disease, ailment orcondition, such as for example, acute macular neuroretinopathy; Behcet'sdisease; choroidal neovascularization; diabetic uveitis; histoplasmosis;infections, such as fungal or viral-caused infections; maculardegeneration, such as acute macular degeneration, non-exudative agerelated macular degeneration and exudative age related maculardegeneration; edema, such as macular edema, cystoid macular edema anddiabetic macular edema; multifocal choroiditis; ocular trauma whichaffects a posterior ocular site or location; ocular tumors; retinaldisorders, such as central retinal vein occlusion, diabetic retinopathy(including proliferative diabetic retinopathy), proliferativevitreoretinopathy (PVR), retinal arterial occlusive disease, retinaldetachment, uveitic retinal disease; sympathetic ophthalmia; VogtKoyanagi-Harada (VKH) syndrome; uveal diffusion; a posterior ocularcondition caused by or influenced by an ocular laser treatment;posterior ocular conditions caused by or influenced by a photodynamictherapy, photocoagulation, radiation retinopathy, epiretinal membranedisorders, branch retinal vein occlusion, anterior ischemic opticneuropathy, non-retinopathy diabetic retinal dysfunction, retinitispigmentosa, and glaucoma. Glaucoma can be considered a posterior ocularcondition because the therapeutic goal is to prevent the loss of orreduce the occurrence of loss of vision due to damage to or loss ofretinal cells or optic nerve cells (i.e. neuroprotection).

“Biodegradable polymer” means a polymer or polymers which degrade invivo, and wherein erosion of the polymer or polymers over time is occursconcurrent with or subsequent to release of the therapeutic agent.Specifically, hydrogels such as methylcellulose which act to releasedrug through polymer swelling are specifically excluded from the term“biodegradable polymer”. The terms “biodegradable” and “bioerodible” areequivalent and are used interchangeably herein. A biodegradable polymermay be a homopolymer, a copolymer, or a polymer comprising more than twodifferent polymeric units.

“Treat”, “treating”, or “treatment” means a reduction or resolution orprevention of an ocular condition, ocular injury or damage, or topromote healing of injured or damaged ocular tissue.

“Therapeutically effective amount” means the level or amount of agentneeded to treat an ocular condition, or reduce or prevent ocular injuryor damage without causing significant negative or adverse side effectsto the eye or a region of the eye.

Intraocular implants have been developed which can release drug loadsover various' time periods. These implants, which when inserted into aneye, such as the vitreous of an eye, provide therapeutic levels of analpha-2 adrenergic receptor agonist for extended periods of time (e.g.,for about 1 week or more). The implants disclosed are effective intreating ocular conditions, such as posterior ocular conditions.

In one embodiment of the present invention, an intraocular implantcomprises a biodegradable polymer matrix. The biodegradable polymermatrix is one type of a drug release sustaining component. Thebiodegradable polymer matrix is effective in forming a biodegradableintraocular implant. The biodegradable intraocular implant comprises analpha-2 adrenergic receptor agonist associated with the biodegradablepolymer matrix. The matrix degrades at a rate effective to sustainrelease of an amount of the alpha-2 adrenergic receptor agonist for atime greater than about one week from the time in which the implant isplaced in ocular region or ocular site, such as the vitreous of an eye.

The alpha-2 adrenergic receptor agonist of the implant is typically anagent that selectively activates alpha-2 adrenergic receptors relativeto alpha-1 adrenergic receptors. In certain implants, the alpha-2adrenergic receptor agonist is selectively activates a subtype of thealpha-2 adrenergic receptors. For example, the agonist may selectivelyactivate one or more of the alpha-2a, the alpha-2b, or the alpha-2creceptors, under certain conditions, such as physiological conditions.Under other conditions, the agonist of the implant may not be selectivefor alpha-2 adrenergic receptor subtypes. The agonist may activate thereceptors by binding to the receptors, or by any other mechanism.

In certain implants, the alpha-2 adrenergic receptor agonist is aquinoxaline derivative. The quinoxaline derivatives useful in thepresent implants are those quinoxaline derivatives having the formula,

pharmaceutically acceptable acid addition salts thereof, and mixturesthereof. R₁ and R₂ each is independently selected from the groupconsisting of H, alkyl radicals containing 1 to 4 carbon atoms andalkoxy radicals containing 1 to 4 carbon atoms. R₂ is preferably amethyl radical. The 2-imidazolin-2-ylamino group may be in any of the5-, 6-, 7- and 8-positions, preferably in the 6-position, of thequinoxaline nucleus. R₃, R₄ and R₅ each is located in one of theremaining 5-, 6-, 7- or 8-positions of the quinoxaline nucleus and isindependently selected from the group consisting of Cl, Br, H and alkylradicals containing 1 to 3 carbon atoms. R₃ is preferably in the5-position of the quinoxaline nucleus, and R₄ and R₅ are preferably bothH. In a particularly useful embodiment R₃ is Br.

In at least one implant, R₁ is H and R₂ is selected from alkyl radicalscontaining 1 to 4 carbon atoms. R₃ may advantageously be in the5-position of the quinoxaline nucleus and be selected from H and alkylradicals containing 1 to 3 carbon atoms. All stereoisomers, tautomersand mixtures thereof which comply with the constraints of one or more ofthe presently useful compounds are included within the scope of thepresent invention.

Pharmaceutically acceptable acid addition salts of the compounds of theinvention are those formed from acids which form non-toxic additionsalts containing pharmaceutically acceptable anions, such as thehydrochloride, hydrobromide, hydroiodide, sulfate, or bisulfate,phosphate or acid phosphate, acetate, maleate, fumarate, oxalate,lactate, tartrate, citrate, gluconate, saccharate and p-toluenesulphonate salts.

In more specific implants, the quinoxaline derivative has the formula

In additional implants, the alpha-2 adrenergic receptor agonist isprovided as a salt having the formula

The foregoing salt is known as brimonidine tartrate (AGN 190342-F,5-bromo-6-(2-imidazolidinylideneamino) quinoxaline tartrate), and ispublicly available from Allergan, Inc. under the tradename Alphagan-P®.Brimonidine, an organic base, is publicly available as eitherbrimonidine tartrate salt or as brimonidine freebase. The tartrate saltis more soluble than the freebase in various aqueous media. Since boththe tartrate salt and the freebase are chemically stable and havemelting points higher than 200° C., both forms are suitable in formingthe present implants.

Thus, the implant may comprise a therapeutic component which comprises,consists essentially of, or consists of a brimonidine salt, such asbrimonidine tartrate, a brimonidine free base, or mixtures thereof.

The alpha-2 adrenergic receptor agonist may be in a particulate orpowder form and entrapped by the biodegradable polymer matrix. Usually,alpha-2 adrenergic receptor agonist particles will have an effectiveaverage size less than about 3000 nanometers. In certain implants, theparticles may have an effective average particle size about an order ofmagnitude smaller than 3000 nanometers. For example, the particles mayhave an effective average particle size of less than about 500nanometers. In additional implants, the particles may have an effectiveaverage particle size of less than about 400 nanometers, and in stillfurther embodiments, a size less than about 200 nanometers.

The alpha-2 adrenergic receptor agonist of the implant is preferablyfrom about 10% to 90% by weight of the implant. More preferably, thealpha-2 adrenergic receptor agonist is from about 20% to about 80% byweight of the implant. In a preferred embodiment, the alpha-2 adrenergicreceptor agonist comprises about 20% by weight of the implant (e.g.,15%-25%). In another embodiment, the alpha-2 adrenergic receptor agonistcomprises about 50% by weight of the implant.

Suitable polymeric materials or compositions for use in the implantinclude those materials which are compatible, that is biocompatible,with the eye so as to cause no substantial interference with thefunctioning or physiology of the eye. Such materials preferably are atleast partially and more preferably substantially completelybiodegradable or bioerodible.

Examples of useful polymeric materials include, without limitation, suchmaterials derived from and/or including organic esters and organicethers, which when degraded result in physiologically acceptabledegradation products, including the monomers. Also, polymeric materialsderived from and/or including, anhydrides, amides, orthoesters and thelike, by themselves or in combination with other monomers, may also finduse. The polymeric materials may be addition or condensation polymers,advantageously condensation polymers. The polymeric materials may becross-linked or non-cross-linked, for example not more than lightlycross-linked, such as less than about 5%, or less than about 1% of thepolymeric material being cross-linked. For the most part, besides carbonand hydrogen, the polymers will include at least one of oxygen andnitrogen, advantageously oxygen. The oxygen may be present as oxy, e.g.hydroxy or ether, carbonyl, e.g. non-oxo-carbonyl, such as carboxylicacid ester, and the like. The nitrogen may be present as amide, cyanoand amino. The polymers set forth in Heller, Biodegradable Polymers inControlled Drug Delivery, In: CRC Critical Reviews in Therapeutic DrugCarrier Systems, Vol. 1, CRC Press, Boca Raton, Fla. 1987, pp 39-90,which describes encapsulation for controlled drug delivery, may find usein the present implants.

Of additional interest are polymers of hydroxyaliphatic carboxylicacids, either homopolymers or copolymers, and polysaccharides.Polyesters of interest include polymers of D-lactic acid, L-lactic acid,racemic lactic acid, glycolic acid, polycaprolactone, and combinationsthereof. Generally, by employing the L-lactate or D-lactate, a slowlyeroding polymer or polymeric material is achieved, while erosion issubstantially enhanced with the lactate racemate.

Among the useful polysaccharides are, without limitation, calciumalginate, and functionalized celluloses, particularlycarboxymethylcellulose esters characterized by being water insoluble, amolecular weight of about 5 kD to 500 kD, for example.

Other polymers of interest include, without limitation, polyvinylalcohol, polyesters, polyethers and combinations thereof which arebiocompatible and may be biodegradable and/or bioerodible.

Some preferred characteristics of the polymers or polymeric materialsfor use in the present invention may include biocompatibility,compatibility with the therapeutic component, ease of use of the polymerin making the drug delivery systems of the present invention, ahalf-life in the physiological environment of at least about 6 hours,preferably greater than about one day, not significantly increasing theviscosity of the vitreous, and water insolubility.

The biodegradable polymeric materials which are included to form thematrix are desirably subject to enzymatic or hydrolytic instability.Water soluble polymers may be cross-linked with hydrolytic orbiodegradable unstable cross-links to provide useful water insolublepolymers. The degree of stability can be varied widely, depending uponthe choice of monomer, whether a homopolymer or copolymer is employed,employing mixtures of polymers, and whether the polymer includesterminal acid groups.

Equally important to controlling the biodegradation of the polymer andhence the extended release profile of the implant is the relativeaverage molecular weight of the polymeric composition employed in theimplant. Different molecular weights of the same or different polymericcompositions may be included in the implant to modulate the releaseprofile. In certain implants, the relative average molecular weight ofthe polymer will range from about 9 to about 64 kD, usually from about10 to about 54 kD, and more usually from about 12 to about 45 kD.

In some implants, copolymers of glycolic acid and lactic acid are used,where the rate of biodegradation is controlled by the ratio of glycolicacid to lactic acid. The most rapidly degraded copolymer has roughlyequal amounts of glycolic acid and lactic acid. Homopolymers, orcopolymers having ratios other than equal, are more resistant todegradation. The ratio of glycolic acid to lactic acid will also affectthe brittleness of the implant, where a more flexible implant isdesirable for larger geometries. The % of polylactic acid in thepolylactic acid polyglycolic acid (PLGA) copolymer can be 0-100%,preferably about 15-85%, more preferably about 35-65%. In some implants,a 50/50 PLGA copolymer is used.

The biodegradable polymer matrix of the intraocular implant may comprisea mixture of two or more biodegradable polymers. For example, theimplant may comprise a mixture of a first biodegradable polymer and adifferent second biodegradable polymer. One or more of the biodegradablepolymers may have terminal acid groups.

Release of a drug from an erodible polymer is the consequence of severalmechanisms or combinations of mechanisms. Some of these mechanismsinclude desorption from the implants surface, dissolution, diffusionthrough porous channels of the hydrated polymer and erosion. Erosion canbe bulk or surface or combination of both. As discussed herein, thematrix of the intraocular implant may release drug at a rate effectiveto sustain release of an amount of the alpha-2 adrenergic receptoragonist for more than one week after implantation into an eye. Incertain implants, therapeutic amounts of the alpha-2 adrenergic receptoragonist are released for no more than about 30-35 days afterimplantation. For example, an implant may comprise brimonidine tartrate,and the matrix of the implant degrades at a rate effective to sustainrelease of a therapeutically effective amount of brimonidine tartratefor about one month after being placed in an eye. As another example,the implant may comprise brimonidine tartrate, and the matrix releasesdrug at a rate effective to sustain release of a therapeuticallyeffective amount of brimonidine tartrate for more than forty days, suchas for about six months.

One example of the biodegradable intraocular implant comprises analpha-2 adrenergic receptor agonist associated with a biodegradablepolymer matrix, which comprises a mixture of different biodegradablepolymers. At least one of the biodegradable polymers is a polylactidehaving a molecular weight of about 63.3 kD. A second biodegradablepolymer is a polylactide having a molecular weight of about 14 kD. Sucha mixture is effective in sustaining release of a therapeuticallyeffective amount of the alpha-2 adrenergic receptor agonist for a timeperiod greater than about one month from the time the implant is placedin an eye.

Another example of a biodegradable intraocular implant comprises analpha-2 adrenergic receptor agonist associated with a biodegradablepolymer matrix, which comprises a mixture of different biodegradablepolymers, each biodegradable polymer having an inherent viscosity fromabout 0.16 dl/g to about 1.0 dl/g. For example, one of the biodegradablepolymers may have an inherent viscosity of about 0.3 dl/g. A secondbiodegradable polymer may have an inherent viscosity of about 1.0 dl/g.The inherent viscosities identified above may be determined in 0.1%chloroform at 25° C.

One particular implant comprises brimonidine tartrate associated with acombination of two different polylactide polymers. The brimonidinetartrate is present in about 20% by weight of the implant. Onepolylactide polymer has a molecular weight of about 14 kD and aninherent viscosity of about 0.3 dl/g, and the other polylactide polymerhas a molecular weight of about 63.3 kD and an inherent viscosity ofabout 1.0 dl/g. The two polylactide polymers are present in the implantin a 1:1 ratio. Such an implant provides for release of the brimonidinefor more than two months in vitro, as described herein. The implant isprovided in the form of a rod or a filament produced by an extrusionprocess.

The release of the alpha-2 adrenergic receptor agonist from theintraocular implant comprising a biodegradable polymer matrix mayinclude an initial burst of release followed by a gradual increase inthe amount of the alpha-2 adrenergic receptor agonist released, or therelease may include an initial delay in release of the alpha-2adrenergic receptor agonist followed by an increase in release. When theimplant is substantially completely degraded, the percent of the alpha-2adrenergic receptor agonist that has been released is about one hundred.Compared to existing implants, the implants disclosed herein do notcompletely release, or release about 100% of the alpha-2 adrenergicreceptor agonist, until after about one week of being placed in an eye.

It may be desirable to provide a relatively constant rate of release ofthe alpha-2 adrenergic receptor agonist from the implant over the lifeof the implant. For example, it may be desirable for the alpha-2adrenergic receptor agonist to be released in amounts from about 0.01 μgto about 2 μg per day for the life of the implant. However, the releaserate may change to either increase or decrease depending on theformulation of the biodegradable polymer matrix. In addition, therelease profile of the alpha-2 adrenergic receptor agonist may includeone or more linear portions and/or one or more non-linear portions.Preferably, the release rate is greater than zero once the implant hasbegun to degrade or erode.

The implants may be monolithic, i.e. having the active agent or agentshomogenously distributed through the polymeric matrix, or encapsulated,where a reservoir of active agent is encapsulated by the polymericmatrix or as a core-shell type of implant. Due to ease of manufacture,monolithic implants are usually preferred over encapsulated forms.However, the greater control afforded by the encapsulated,reservoir-type implant may be of benefit in some circumstances, wherethe therapeutic level of the drug falls within a narrow window. Inaddition, the therapeutic component, including the alpha-2 adrenergicreceptor agonist, may be distributed in a non-homogenous pattern in thematrix. For example, the implant may include a portion that has agreater concentration of the alpha-2 adrenergic receptor agonistrelative to a second portion of the implant.

The intraocular implants disclosed herein may have a size of betweenabout 5 μm and about 2 mm, or between about 10 μm and about 1 mm foradministration with a needle, greater than 1 mm, or greater than 2 mm,such as 3 mm or up to 10 mm, for administration by surgicalimplantation. The vitreous chamber in humans is able to accommodaterelatively large implants of varying geometries, having lengths of, forexample, 1 to 10 mm. The implant may be a cylindrical pellet (e.g., rod)with dimensions of about 2 mm×0.75 mm diameter. Or the implant may be acylindrical pellet with a length of about 7 mm to about 10 mm, and adiameter of about 0.75 mm to about 1.5 mm.

The implants may also be at least somewhat flexible so as to facilitateboth insertion of the implant in the eye, such as in the vitreous, andaccommodation of the implant. The total weight of the implant is usuallyabout 250-5000 μg, more preferably about 500-1000 μg. For example, animplant may be about 500 μg, or about 1000 μg. For non-humanindividuals, the dimensions and total weight of the implant(s) may belarger or smaller, depending on the type of individual. For example,humans have a vitreous volume of approximately 3.8 ml, compared withapproximately 30 ml for horses, and approximately 60-100 ml forelephants. An implant sized for use in a human may be scaled up or downaccordingly for other animals, for example, about 8 times larger for animplant for a horse, or about, for example, 26 times larger for animplant for an elephant.

Thus, implants can be prepared where the center may be of one materialand the surface may have one or more layers of the same or a differentcomposition, where the layers may be cross-linked, or of a differentmolecular weight, different density or porosity, or the like. Forexample, where it is desirable to quickly release an initial bolus ofdrug, the center may be a polylactate coated with apolylactate-polyglycolate copolymer, so as to enhance the rate ofinitial degradation. Alternatively, the center may be polyvinyl alcoholcoated with polylactate, so that upon degradation of the polylactateexterior the center would dissolve and be rapidly washed out of the eye.

The implants may be of any geometry including fibers, sheets, films,microspheres, spheres, circular discs, plaques and the like. The upperlimit for the implant size will be determined by factors such astoleration for the implant, size limitations on insertion, ease ofhandling, etc. Where sheets or films are employed, the sheets or filmswill be in the range of at least about 0.5 mm×0.5 mm, usually about 3-10mm×5-10 mm with a thickness of about 0.1-1.0 mm for ease of handling.Where fibers are employed, the fiber diameter will generally be in therange of about 0.05 to 3 mm and the fiber length will generally be inthe range of about 0.5-10 mm. Spheres may be in the range of 0.5 μm to 4mm in diameter, with comparable volumes for other shaped particles.

The size and form of the implant can also be used to control the rate ofrelease, period of treatment, and drug concentration at the site ofimplantation. Larger implants will deliver a proportionately largerdose, but depending on the surface to mass ratio, may have a slowerrelease rate. The particular size and geometry of the implant are chosento suit the site of implantation.

The proportions of alpha-2 adrenergic receptor agonist, polymer, and anyother modifiers may be empirically determined by formulating severalimplants with varying proportions. A USP approved method for dissolutionor release test can be used to measure the rate of release (USP 23; NF18 (1995) pp. 1790-1798). For example, using the infinite sink method, aweighed sample of the implant is added to a measured volume of asolution containing 0.9% NaCl in water, where the solution volume willbe such that the drug concentration is after release is less than 5% ofsaturation. The mixture is maintained at 37° C. and stirred slowly tomaintain the implants in suspension. The appearance of the dissolveddrug as a function of time may be followed by various methods known inthe art, such as spectrophotometrically, HPLC, mass spectroscopy, etc.until the absorbance becomes constant or until greater than 90% of thedrug has been released.

In addition to the alpha-2 adrenergic receptor agonist or alpha-2adrenergic receptor agonists included in the intraocular implantsdisclosed herein, the intraocular implants may also include one or moreadditional ophthalmically acceptable therapeutic agents. For example,the implant may include one or more antihistamines, one or moreantibiotics, one or more beta blockers, one or more steroids, one ormore antineoplastic agents, one or more immunosuppressive agents, one ormore antiviral agents, one or more antioxidant agents, and mixturesthereof.

Pharmacologic or therapeutic agents which may find use in the presentsystems, include, without limitation, those disclosed in U.S. Pat. Nos.4,474,451, columns 4-6 and 4,327,725, columns 7-8.

Examples of antihistamines include, and are not limited to, loratadine,hydroxyzine, diphenhydramine, chlorpheniramine, brompheniramine,cyproheptadine, terfenadine, clemastine, triprolidine, carbinoxamine,diphenylpyraline, phenindamine, azatadine, tripelennamine,dexchlorpheniramine, dexbrompheniramine, methdilazine, and trimeprazinedoxylamine, pheniramine, pyrilamine, chlorcyclizine, thonzylamine, andderivatives thereof.

Examples of antibiotics include without limitation, cefazolin,cephradine, cefaclor, cephapirin, ceftizoxime, cefoperazone, cefotetan,cefutoxime, cefotaxime, cefadroxil, ceftazidime, cephalexin,cephalothin, cefamandole, cefoxitin, cefonicid, ceforanide, ceftriaxone,cefadroxil, cephradine, cefuroxime, ampicillin, amoxicillin,cyclacillin, ampicillin, penicillin G, penicillin V potassium,piperacillin, oxacillin, bacampicillin, cloxacillin, ticarcillin,azlocillin, carbenicillin, methicillin, nafcillin, erythromycin,tetracycline, doxycycline, minocycline, aztreonam, chloramphenicol,ciprofloxacin hydrochloride, clindamycin, metronidazole, gentamicin,lincomycin, tobramycin, vancomycin, polymyxin B sulfate, colistimethate,colistin, azithromycin, augmentin, sulfamethoxazole, trimethoprim, andderivatives thereof.

Examples of beta blockers include acebutolol, atenolol, labetalol,metoprolol, propranolol, timolol, and derivatives thereof.

Examples of steroids include corticosteroids, such as cortisone,prednisolone, fluorometholone, dexamethasone, medrysone, loteprednol,fluazacort, hydrocortisone, prednisone, betamethasone, prednisone,methylprednisolone, triamcinolone hexacetonide, paramethasone acetate,diflorasone, fluocinonide, fluocinolone, triamcinolone, derivativesthereof, and mixtures thereof.

Examples of antineoplastic agents include adriamycin, cyclophosphamide,actinomycin, bleomycin, daunorubicin, doxorubicin, epirubicin,mitomycin, methotrexate, fluorouracil, carboplatin, carmustine (BCNU),methyl-CCNU, cisplatin, etoposide, interferons, camptothecin andderivatives thereof, phenesterine, taxol and derivatives thereof,taxotere and derivatives thereof, vinblastine, vincristine, tamoxifen,etoposide, piposulfan, cyclophosphamide, and flutamide, and derivativesthereof.

Examples of immunosuppressive agents include cyclosporine, azathioprine,tacrolimus, and derivatives thereof.

Examples of antiviral agents include interferon gamma, zidovudine,amantadine hydrochloride, ribavirin, acyclovir, valaciclovir,dideoxycytidine, phosphonoformic acid, ganciclovir, and derivativesthereof.

Examples of antioxidant agents include ascorbate, alpha-tocopherol,mannitol, reduced glutathione, various carotenoids, cysteine, uric acid,taurine, tyrosine, superoxide dismutase, lutein, zeaxanthin,cryptpxanthin, astaxanthin, lycopene, N-acetyl-cysteine, carnosine,gamma-glutamylcysteine, quercetin, lactoferrin, dihydrolipoic acid,citrate, Ginkgo Biloba extract, tea catechins, bilberry extract,vitamins E or esters of vitamin E, retinyl palmitate, and derivativesthereof.

Other therapeutic agents include squalamine, carbonic anhydraseinhibitors, alpha agonists, prostamides, prostaglandins, antiparasitics,antifungals, and derivatives thereof.

The amount of active agent or agents employed in the implant,individually or in combination, will vary widely depending on theeffective dosage required and the desired rate of release from theimplant. Usually the agent will be at least about 1, more usually atleast about 10 weight percent of the implant, and usually not more thanabout 80, more usually not more than about 40 weight percent of theimplant.

In addition to the therapeutic component, the intraocular implantsdisclosed herein may include effective amounts of buffering agents,preservatives and the like. Suitable water soluble buffering agentsinclude, without limitation, alkali and alkaline earth carbonates,phosphates, bicarbonates, citrates, borates, acetates, succinates andthe like, such as sodium phosphate, citrate, borate, acetate,bicarbonate, carbonate and the like. These agents advantageously presentin amounts sufficient to maintain a pH of the system of between about 2to about 9 and more preferably about 4 to about 8. As such the bufferingagent may be as much as about 5% by weight of the total implant.Suitable water soluble preservatives include sodium bisulfite, sodiumbisulfate, sodium thiosulfate, ascorbate, benzalkonium chloride,chlorobutanol, thimerosal, phenylmercuric acetate, phenylmercuricborate, phenylmercuric nitrate, parabens, methylparaben, polyvinylalcohol, benzyl alcohol, phenylethanol and the like and mixturesthereof. These agents may be present in amounts of from 0.001 to about5% by weight and preferably 0.01 to about 2% by weight. In at least oneof the present implants, a purite preservative is provided in theimplant, such as when the alpha-2 adrenergic receptor agonist isbrimonidine. Thus, these implants may contain a therapeuticallyeffective amount of Alphagan-P®.

In some situations mixtures of implants may be utilized employing thesame or different pharmacological agents. In this way, a cocktail ofrelease profiles, giving a biphasic or triphasic release with a singleadministration is achieved, where the pattern of release may be greatlyvaried.

Additionally, release modulators such as those described in U.S. Pat.No. 5,869,079 may be included in the implants. The amount of releasemodulator employed will be dependent on the desired release profile, theactivity of the modulator, and on the release profile of the alpha-2adrenergic receptor agonist in the absence of modulator. Electrolytessuch as sodium chloride and potassium chloride may also be included inthe implant. Where the buffering agent or enhancer is hydrophilic, itmay also act as a release accelerator. Hydrophilic additives act toincrease the release rates through faster dissolution of the materialsurrounding the drug particles, which increases the surface area of thedrug exposed, thereby increasing the rate of drug bioerosion. Similarly,a hydrophobic buffering agent or enhancer dissolve more slowly, slowingthe exposure of drug particles, and thereby slowing the rate of drugbioerosion.

In certain implants, an implant comprising brimonidine or brimonidinetartrate and a biodegradable polymer matrix is able to release ordeliver an amount of brimonidine between about 0.1 mg to about 0.5 mgfor about 3-6 months after implantation into the eye. The implant may beconfigured as a rod or a wafer. A rod-shaped implant may be derived fromfilaments extruded from a 720 μm nozzle and cut into 1 mg size. Awafer-shaped implant may be a circular disc having a diameter of about2.5 mm, a thickness of about 0.127 mm, and a weight of about 1 mg.

The proposed 3-month release formulations may be sterile, andbioerodible in the form of a rod, a wafer or a microsphere containingbrimonidine tartrate within a PLA matrix or POE matrix. The implants aredesigned to delay the clearance of the drug and reduce the need forrepeated implantation over 3-month period, thereby lowering the risk ofcomplications.

Various techniques may be employed to produce the implants describedherein. Useful techniques include, but are not necessarily limited to,solvent evaporation methods, phase separation methods, interfacialmethods, molding methods, injection molding methods, extrusion methods,co-extrusion methods, carver press method, die cutting methods, heatcompression, combinations thereof and the like.

Specific methods are discussed in U.S. Pat. No. 4,997,652. Extrusionmethods may be used to avoid the need for solvents in manufacturing.When using extrusion methods, the polymer and drug are chosen so as tobe stable at the temperatures required for manufacturing, usually atleast about 85 degrees Celsius. Extrusion methods use temperatures ofabout 25 degrees C. to about 150 degrees C., more preferably about 65degrees C. to about 130 degrees C. An implant may be produced bybringing the temperature to about 60 degrees C. to about 150 degrees C.for drug/polymer mixing, such as about 130 degrees C., for a time periodof about 0 to 1 hour, 0 to 30 minutes, or 5-15 minutes. For example, atime period may be about 10 minutes, preferably about 0 to 5 min. Theimplants are then extruded at a temperature of about 60 degrees C. toabout 130 degrees C., such as about 75 degrees C.

In addition, the implant may be coextruded so that a coating is formedover a core region during the manufacture of the implant.

Compression methods may be used to make the implants, and typicallyyield implants with faster release rates than extrusion methods.Compression methods may use pressures of about 50-150 psi, morepreferably about 70-80 psi, even more preferably about 76 psi, and usetemperatures of about 0 degrees C. to about 115 degrees C., morepreferably about 25 degrees C.

The implants of the present invention may be inserted into the eye, forexample the vitreous chamber of the eye, by a variety of methods,including placement by forceps or by trocar following making a 2-3 mmincision in the sclera. One example of a device that may be used toinsert the implants into an eye is disclosed in U.S. Patent PublicationNo. 2004/0054374. The method of placement may influence the therapeuticcomponent or drug release kinetics. For example, delivering the implantwith a trocar may result in placement of the implant deeper within thevitreous than placement by forceps, which may result in the implantbeing closer to the edge of the vitreous. The location of the implantmay influence the concentration gradients of therapeutic component ordrug surrounding the element, and thus influence the release rates(e.g., an element placed closer to the edge of the vitreous may resultin a slower release rate).

The present implants are configured to release an amount of alpha-2adrenergic receptor agonist in an eye for a period of time to minimizean ocular vascular occlusion, such as a retinal vascular occlusion.Retinal vascular occlusion may result from a variety of diseases such asretinal arterial occlusive disease, central retinal vein occlusion,disseminated intravascular coagulopathy, branch retinal vein occlusion,hypertensive fundus changes, ocular ischemic syndrome, retinal arterialmicroaneurysms, hemi-retinal vein occlusion, central retinal arteryocclusion, branch retinal artery occlusion, carotid artery disease(cad), eales disease and vasculopathies associated with diabetes. Byimplanting the alpha-2 adrenergic receptor agonist-containing implantsinto the vitreous of an eye, it is believed that the agonist iseffective to reduce occlusion within blood vessels located in the eye.

In addition, the present implants may be configured to release analpha-2 adrenergic receptor agonist in a therapeutically effectiveamount for a period of time effective to treat glaucoma of a patient.

The implants disclosed herein may also be configured to releaseadditional therapeutic agents, as described above, which may beeffective in treating diseases or conditions, such as the following:

Maculopathies/retinal degeneration: macular degeneration, including agerelated macular degeneration (ARMD), such as non-exudative age relatedmacular degeneration and exudative age related macular degeneration,choroidal neovascularization, retinopathy, including diabeticretinopathy, acute and chronic macular neuroretinopathy, central serouschorioretinopathy, and macular edema, including cystoid macular edema,and diabetic macular edema. Uveitis/retinitis/choroiditis: acutemultifocal placoid pigment epitheliopathy, Behcet's disease, birdshotretinochoroidopathy, infectious (syphilis, lyme, tuberculosis,toxoplasmosis), uveitis, including intermediate uveitis (pars planitis)and anterior uveitis, multifocal choroiditis, multiple evanescent whitedot syndrome (MEWDS), ocular sarcoidosis, posterior scleritis,serpiginous choroiditis, subretinal fibrosis, uveitis syndrome, andVogt-Koyanagi-Harada syndrome. Vascular diseases/exudative diseases:retinal arterial occlusive disease, central retinal vein occlusion,disseminated intravascular coagulopathy, branch retinal vein occlusion,hypertensive fundus changes, ocular ischemic syndrome, retinal arterialmicroaneurysms, Coat's disease, parafoveal telangiectasia, hemi-retinalvein occlusion, papillophlebitis, central retinal artery occlusion,branch retinal artery occlusion, carotid artery disease (CAD), frostedbranch angiitis, sickle cell retinopathy and other hemoglobinopathies,angioid streaks, familial exudative vitreoretinopathy, Eales disease.Traumatic/surgical: sympathetic ophthalmia, uveitic retinal disease,retinal detachment, trauma, laser, PDT, photocoagulation, hypoperfusionduring surgery, radiation retinopathy, bone marrow transplantretinopathy. Proliferative disorders: proliferative vitreal retinopathyand epiretinal membranes, proliferative diabetic retinopathy. Infectiousdisorders: ocular histoplasmosis, ocular toxocariasis, presumed ocularhistoplasmosis syndrome (PONS), endophthalmitis, toxoplasmosis, retinaldiseases associated with HIV infection, choroidal disease associatedwith HIV infection, uveitic disease associated with HIV Infection, viralretinitis, acute retinal necrosis, progressive outer retinal necrosis,fungal retinal diseases, ocular syphilis, ocular tuberculosis, diffuseunilateral subacute neuroretinitis, and myiasis. Genetic disorders:retinitis pigmentosa, systemic disorders with associated retinaldystrophies, congenital stationary night blindness, cone dystrophies,Stargardt's disease and fundus flavimaculatus, Bests disease, patterndystrophy of the retinal pigmented epithelium, X-linked retinoschisis,Sorsby's fundus dystrophy, benign concentric maculopathy, Bietti'scrystalline dystrophy, pseudoxanthoma elasticum. Retinal tears/holes:retinal detachment, macular hole, giant retinal tear. Tumors: retinaldisease associated with tumors, congenital hypertrophy of the RPE,posterior uveal melanoma, choroidal hemangioma, choroidal osteoma,choroidal metastasis, combined hamartoma of the retina and retinalpigmented epithelium, retinoblastoma, vasoproliferative tumors of theocular fundus, retinal astrocytoma, intraocular lymphoid tumors.Miscellaneous: punctate inner choroidopathy, acute posterior multifocalplacoid pigment epitheliopathy, myopic retinal degeneration, acuteretinal pigment epithelitis and the like.

In one embodiment, an implant, such as the implants disclosed herein, isadministered to a posterior segment of an eye of a human or animalpatient, and preferably, a living human or animal. In at least oneembodiment, an implant is administered without accessing the subretinalspace of the eye. For example, a method of treating a patient mayinclude placing the implant directly into the posterior chamber of theeye. In other embodiments, a method of treating a patient may compriseadministering an implant to the patient by at least one of intravitrealinjection, subconjunctival injection, sub-tenon injections, retrobulbarinjection, and suprachoroidal injection.

In at least one embodiment, a method of reducing retinal vascularocclusion in a patient comprises administering one or more implantscontaining one or more alpha-2 adrenergic receptor agonists, asdisclosed herein to a patient by at least one of intravitreal injection,subconjunctival injection, sub-tenon injection, retrobulbar injection,and suprachoroidal injection. A syringe apparatus including anappropriately sized needle, for example, a 27 gauge needle or a 30 gaugeneedle, can be effectively used to inject the composition with theposterior segment of an eye of a human or animal. Repeat injections areoften not necessary due to the extended release of the alpha-2adrenergic receptor agonists from the implants.

In another aspect of the invention, kits for treating an ocularcondition of the eye are provided, comprising: a) a container comprisingan extended release implant comprising a therapeutic component includingan alpha-2 adrenergic receptor agonist, such as brimonidine free base orbrimonidine tartrate (e.g., Alphagan-P®), and a drug release sustainingcomponent; and b) instructions for use. Instructions may include stepsof how to handle the implants, how to insert the implants into an ocularregion, and what to expect from using the implants.

Example 1 Manufacture and Testing of Implants Containing Brimonidine anda Biodegradable Polymer Matrix

Biodegradable implants were made by combining brimonidine tartrate orbrimonidine freebase with a biodegradable polymer composition in astainless steel mortar. The combination was mixed via a Turbula shakerset at 96 RPM for 15 minutes. The powder blend was scraped off the wallof the mortar and then remixed for an additional 15 minutes. The mixedpowder blend was heated to a semi-molten state at specified temperaturefor a total of 30 minutes, forming a polymer/drug melt.

Rods were manufactured by pelletizing the polymer/drug melt using a 9gauge polytetrafluoroethylene (PTFE) tubing, loading the pellet into thebarrel and extruding the material at the specified core extrusiontemperature into filaments. The filaments were then cut into about 1 mgsize implants or drug delivery systems. The rods had dimensions of about2 mm long×0.72 mm diameter. The rod implants weighed between about 900μg and 1100 μg.

Wafers were formed by flattening the polymer melt with a Carver press ata specified temperature and cutting the flattened material into wafers,each weighing about 1 mg. The wafers had a diameter of about 2.5 mm anda thickness of about 0.13 mm. The wafer implants weighed between about900 μg and 1100 μg.

The in-vitro release testing was performed on each lot of implant (rodor wafer) in six replicates initially, and later in four replicates.Each implant was placed into a 24 mL screw cap vial with 10 mL ofPhosphate Buffered Saline solution at 37° C. and 1 mL aliquots wereremoved and replaced with equal volume of fresh medium on day 1, 4, 7,14, 28, and every two weeks thereafter.

The drug assays were performed by HPLC, which consists of a Waters 2690Separation Module (or 2696), and a Waters 2996 Photodiode ArrayDetector. An Ultrasphere, C-18 (2), 5 μm; 4.6×150 mm column heated at30° C. was used for separation and the detector was set at 264 nm. Themobile phase was (10:90) MeOH-buffered mobile phase with a flow rate of1 mL/min and a total run time of 12 min per sample. The buffered mobilephase comprised of (68:0.75:0.25:31) 13 mM 1-Heptane Sulfonic Acid,sodium salt-glacial acetic acid-triethylamine-Methanol. The releaserates were determined by calculating the amount of drug being releasedin a given volume of medium over time in μg/day.

The polymers chosen for the implants are were obtained from BoehringerIngelheim. The polymers were: RG502, RG752, R202H, R203 and R206, andPurac PDLG (50/50). RG502 is (50:50) poly(D,L-lactide-co-glycolide),RG752 is (75:25) poly(D,L-lactide-co-glycolide), R202H is 100% poly(D,L-lactide) with acid end group or terminal acid groups, R203 and R206are both 100% poly(D, L-lactide). Purac PDLG (50/50) is (50:50)poly(D,L-lactide-co-glycolide). The inherent viscosity of RG502, RG752,R202H, R203, R206, and Purac PDLG are 0.2, 0.2, 0.2, 0.3, 1.0, and 0.2dL/g, respectively. The average molecular weight of RG502, RG752, R202H,R203, R206, and Purac PDLG are, 11700, 11200, 6500, 14000, 63300, and9700 Daltons, respectively.

A total of 53 formulations were prepared, 31 rods and 22 wafers. Of therod formulations, 4 had release periods longer than 3 months and 3 hadrelease periods longer than 6 months. Of the wafer formulations, 7 hadrelease periods longer than 3 months and 4 had release periods longerthan 4 months.

A list of the rod formulations is shown in Table 1, and a list of waferformulations is shown in Table 2.

TABLE 1 Brimonidine Rod Formulations Formulation Lot BT (w/w) BFB (w/w)Polymer I.V. (dL/g) Core Extr T 1  295-123 50% RG752 0.2 104° C. 2 295-124 50% RG752 0.2 105° C. 3  295-126 50% RG502 0.2 108° C. 4 295-127 50% RG502 0.2 112° C. 5  295-167 50% R203 0.3  98° C. 6 295-168 50% R203 0.3 101° C. 7  295-169 50% R206 1.0 118° C. 8  295-17050% R206 1.0 104° C. 9  295-171 25% R206 1.0  98° C. 10  295-172 25%R203 0.3  96° C. 11 453-3  10% 40% R203 0.3  98° C. 12 453-4  5% 20%R203 0.3  96° C. 13 453-6  10% 40% R206 1.0 105° C. 14 453-7  5% 20%R206 1.0 104° C. 15 453-8  5% 45% R206 1.0 102° C. 16 453-9  15% R2061.0 102° C. 17 453-10 20% (1:1) R203/R206 N/A  98° C. 18 453-11 20%(3:1) R203/R206 N/A  96° C. 19 453-12 10% 40% RG752 0.2 108° C. 20453-13 5% 20% RG752 0.2 104° C. 24 453-50 20% R206 1.0 100° C. 25 453-5117% (1:1) R203/R206 N/A  98° C. 26 453-52 40% (1:1) RG752/RG502 N/A 105°C. 27 453-53 40% (3:1) RG752/RG502 N/A 103° C. 28 453-54 40% (1:1)R203/RG502 N/A 103° C. 29 453-55 50% R202H 0.2  96° C. 30 453-56 50%R202H 0.2  98° C. 31 453-73 20% RG752 0.2  98° C. 32 453-74 20% Purac(Mw 9700) N/A  95° C. 33 453-75 20% Purac (Mw 9700) N/A  92° C. 53453-95 20% (2:1) R203/R206 N/A  97° C. BT = Brimonidine Tartrate BFB =Brimonidine Free Base I.V. = Inherent Viscosity

TABLE 2 Brimonidine wafer Formulations BT BFB I.V. Formulation Lot (w/w)(w/w) Polymer (dL/g) 21 453-47 25% R206 1.0 22 453-48 20% (1:1)R203/R206 N/A 23 453-49 20% (3:1) R203/R206 N/A 34 453-76 20% (1:1)R203/R206 N/A 35 453-77 25% R206 1.0 36 453-78 20% (3:1) R203/R206 N/A37 453-79 25% R203 0.3 38 453-80 50% R203 0.3 39 453-81 50% R206 1.0 40453-82 15% R206 1.0 41 453-83 40% (1:1) RG752/RG502 N/A 42 453-84 40%(2:1) RG752/RG502 N/A 43 453-85 40% (1:1) R203/RG502 N/A 44 453-86 50%R202H 0.2 45 453-87 50% (1:1) RG752/RG502 N/A 46 453-88 10% (1:1)R203/R206 N/A 47 453-89 15% (1:1) R203/R206 N/A 48 453-90 10% (3:1)R203/R206 N/A 49 453-91 15% (3:1) R203/R206 N/A 50 453-92 10% R206 1.051 453-93 10% (2:1) R203/R206 N/A 52 453-94 15% (2:1) R203/R206 N/A

BT=Brimonidine Tartrate BFB=Brimonidine Free Base I.V.=InherentViscosity Rod Formulations

The first 10 formulations were prepared with the five differentpolymers, RG752, RG502, R203, R206, and R202H each at 50% w/w drug loadfor both brimonidine tartrate and brimonidine free base. The releaseprofiles are shown in FIG. 1 for brimonidine tartrate and FIG. 2 forbrimonidine free base.

In most cases, formulations prepared with brimonidine tartrate had afaster initial burst than those prepared from brimonidine freebase usingthe same polymer, except for RG502. The data also show that brimonidinefreebase had a lag time of approximately 30 days when formulated inpoly(D, L-lactide) matrix (R203, R206, and R202H), while brimonidinetartrate was released completely on the first day (F5 and F7). This maybe due to the quick dissolution of brimonidine tartrate on the surfaceof the implant.

Several formulations using R203 and R206 with drug doses lower than 50%were prepared, and the release profiles are shown in FIG. 3. Dramaticeffects were observed when the drug load was lowered from 50% down to25%. For example, formulation #9 was prepared with 25% brimonidinetartrate in R206 and it gave a total release of 89% after 105 daysbefore leveling off. Comparing this to formulation #7, which was 50%brimonidine tartrate in R206, and it released 100% in one day.Similarly, formulation #10 was prepared with 25% brimonidine tartrate inR203 and it gave a total release of 90% after 105 days before it leveledoff. Comparing this to formulation #5, which released 74% on day one.

With 20% brimonidine tartrate in R206 (F24), a 14 day lag time ispresent before it started releasing and eventually reaching 89.5%release after 134 days. At 15% brimonidine tartrate in R206 (F16), thelag time increased to 28 days before it started releasing and eventuallyreaching 97.6% after 175 days.

The release profiles of formulation #9 and #10 behaved in an oppositebut complementary way, in that one polymer exhibits early release whilethe other exhibits a delayed release, but both reached the same endpoint at the same time. When both polymers were combined with a lowerdrug load, a more linear and longer release profile would be obtained,as shown in FIG. 4.

The data show that formulation #17, 20% brimonidine tartrate/(1:1)R203/R206, has a desirable in-vitro release profile for a six monthrelease implant. It released approximately 90% of the brimonidinetartrate after 175 days. It was also shown that by varying theproportion of R203 and R206, even with the same drug load (Formulation#17, #18, and #53), different release profiles would result.

Brimonidine freebase formulations with polymer blends were also preparedto see if a more linear release profile could be obtained. Knowing itslow solubility in aqueous media and its release characteristics in eachpolymer, different combinations of RG502-RG752, and RG502-R203 wereprepared, and the release profiles are shown in FIG. 5.

The duration of release for all three formulations was approximately 2months, but all three exhibited a lag time between 1 to 2 weeks. Twoformulations (F32 and F33) were prepared with Purac polymer, PDLG(50/50)-Mw 9700, one with brimonidine tartrate and the one withbrimonidine freebase. Both formulations had fast release with highstandard of deviation; therefore, the release tests were stopped after 7days.

Wafer Formulations

The first set of wafer formulations was prepared from 3 existing rodformulations. Specifically, formulations #9, #17 and #18, with releasereaching 89.4% after 105 days, 89.2% after 175 days, and 102% after 175days, respectively. The release profiles of the first three waferformulations are shown in FIG. 6.

These three formulations had release periods lasting only two to threeweeks, while their rod counterparts had release periods lasting three tofour months. This may be due to the increased surface area of the wafercompared to that of a rod. In the wafer configuration, drug load alsodetermines the duration of drug release. Therefore, drug load wasreduced from 20-25% down to 15% and 10% and the release profiles areshown in FIGS. 7 and 8.

At 15% drug load, formulation #7 had a cumulative release 51.4% after 35days, while formulation #47, 49, and 52 had cumulative releases of93.2%, 92.8% and 88.5%, respectively, after 99 days. The latter threeformulations may be effective as a 4-month drug delivery system.

At 10% drug load, formulations #46, #48, #50, and #51 had cumulativereleases of 83.8%, 98.0%, 92.7% and 89.2%, respectively, after 133 days.These four formulations may be effective as 5-month drug deliverysystems. Both FIGS. 7 and 8 demonstrate that lowering the drug loadyielded not only a longer duration of release but also more linearrelease profiles for all formulations. The figures also show that usinga polymer blend instead of just a single polymer, such as R206, shouldyield a more linear release profile with lower standard of deviations.

Three wafer formulations were prepared from three previous rodformulations #26, #27, and #28, and the release profiles are shown inFIG. 9. The three wafer formulations released slightly faster than theirrod counterparts at day 28 and they were expected to complete theirrelease between days 31 to 55.

Conclusions

Of the 15 rod formulations prepared from brimonidine tartrate, threeformulations had release periods longer than 3 months (F9, F10, andF53), two formulations had release periods longer than 4 months (F24 andF25), and three formulations had release periods close to 6 months (F16,F17, and F18). Of the 8 rod formulations prepared from brimonidinefreebase, 3 had release periods longer than 2 months (F26, F27, andF28).

Of the 22 wafer formulations, 11 were prepared from brimonidine tartrateand 11 were prepared from brimonidine freebase. Of the 11 waferformulations prepared from brimonidine tartrate, 3 had release periodsof about 4 months (F47, F49, and F52), and 4 had release periods between4 and 5 months (F46, F48, F50, and F51). Of the 11 wafer formulationsprepared from brimonidine freebase, 4 had release periods between 3 and4 months (F35, F36, F38, and F39), and 5 had release periods between oneto two months (F34, F37, F41, F42, and F43).

In general, the wafer formulations prepared from brimonidine tartrate orbrimonidine freebase have faster release than their rod counterparts.

Example 2 In Vivo Testing of Intraocular Implants Containing Brimonidineand a Biodegradable Polymer Matrix

Cynomolgous monkeys were randomly assigned to receive either placebo(n=2) or brimonidine (n=2) formulated intravitreal implants. Baselinemeasures were performed 3 days prior to implantation and 10 daysfollowing implantation with intraocular pressure (IOP), mfERG, laserDoppler scanning topography/flowmetry (HRT/HRF), optical coherencetomography (OCT), indocyanine green angiography (ICG) and fluoresceinangiography (FA).

Three implants (Formulation #17 described in Example 1), each formulatedwith 200 μg brimonidine or placebo were implanted intravitreally into aneye through a port made with an MVR blade (OS), the port was closed withsutures. Wide angle contact lens fundus photography verified implantcount and localization.

Branch retinal vein occlusion (BRVO) was achieved by injecting 1 ml of20 mg/kg Rose Bengal intravenously followed by thermal irradiation usingOmni Coherent Diode laser at 532 nm, 600 mW, 50 um spot size, 0.01 secpulse mode with a 1.6× inversion contact lens. Laser pulses weredelivered until the vein segment was closed. One brimonidine treatedmonkey received 235 pulses and the other received 78 pulses. One placebotreated monkey received 43 pulses and the other received 31 pulses.Vascular occlusion of a vein was induced in the superior arcadeapproximately one disc diameter from the optic nerve head. Occlusion wasverified post-laser by fundus photography.

Funduscopic observations at day 1 following BRVO showed dramaticretinopathy and vasculopathy in both monkeys with placebo implant—markedretinal edema and dot blot hemorrhages, vessel tortuosity, cotton woolspots. Fluorescein angiography verified vein occlusion and stagnateblood flow upstream from the lasered region and elucidated late phasefluorescein leak and pooling from retina capillaries. Monkeys withbrimonidine implants had less than 5 small dot blot hemorrhages, someretinal edema localized to the superior retina. Fluorescein angiographyin brimonidine monkeys showed reperfusion of the once occluded vein withminimal stagnate blood flow.

The brimonidine containing implants decreased the duration of vascularocclusion as shown in FIG. 10. Delay in fluorescein filling of theoccluded vein was quantified using Metamorph 6.0 software. Intensitymeasurements were made with pre-defined regions of interest for earlyand late phases of fluorescein angiography to quantify delay in fillingand the observed delay in fluorescein clearance. The delay in earlyphase filling of fluorescein (seconds) in the occluded vein frombaseline fluorescein angiography filling is illustrated in FIG. 10.

Fovea thickness measurements from OCT single line scans (6 mm) show anincrease in retinal edema as a result of vascular occlusion in theplacebo group. Brimonidine containing implants decreased the magnitudeof retinal edema associated with vascular occlusion. A series of linescans (covering 3 mm²) directly compare changes in retinal thickness inthe superior region surrounding the occluded vein with thickness changesin the inferior retina. Retinal edema in placebo monkeys was so profoundthat fluid accumulation occurred in the inferior region of the retina.In contrast, the brimonidine group did not have a significant change ininferior retina edema compared to baseline, as shown in FIG. 11.

Intraocular pressure (IOP) was recorded (OD and OS) in each group intriplicate post implantation and prior to all follow-upelectrophysiology and retinal imaging procedures. The brimonidineimplants did not significantly lower IOP in eyes prior to or duringBRVO, as shown in FIG. 12

Multi-focal ERG was performed using a VERIS 5.0 system. A stimulus fieldof 241 hexagons was positioned to record superior retina and centralretina foveal response. In the placebo group, foveal responses wereabsent through 3-4 weeks post BRVO induction, whereas, the fovealresponse in the brimonidine group was slightly lower but pronounced atday 1 following BRVO, with recovery and/or higher foveal response forthe remainder of the study. The graph in FIG. 13 shows thesuperior/inferior % response for both groups. BRVO in monkeys treatedwith placebo have less responsive retinal function with a trend towardrecovery late in the study versus relatively consistent retinal functionwith brimonidine implants.

Laser Doppler Flowmetry (HRF) was used to measure blood flow in thefovea, superior and inferior retina regions. The graph of FIG. 14 showsthe results from blood flow measurements acquired with a 10-20 degreezone, centered at the fovea. Blood flow in the fovea appears to beunchanged in the brimonidine group following BRVO, but is sharplyelevated at day 1 post BRVO in the placebo group.

Intravitreal application of three brimonidine intraocular implants haslessened the magnitude and duration of localized vascular occlusion andassociated vasculopathy and retinopathy in monkeys.

In addition, the amount of laser burns needed to close the veins washigher in the brimonidine group compared to placebo (brimonidine:157±79, n=2; placebo: 37±6, n=2). Together, these data show that thepresence of brimonidine increases the difficulty of occluding retinalvasculature and decreases the duration of that occlusion.

Example 3 Treatment of Glaucoma with an Intraocular Implant ContainingBrimonidine Associated with a Biodegradable Polymer Matrix

A 68 year old female complains to her physician that it is becomingdifficult to see. The physician determines that she has elevatedintraocular pressure levels, and diagnoses her with glaucoma. An implantcontaining 200 μg of brimonidine tartrate and 800 μg of a combination ofbiodegradable polymers (R203 and R206 at a 1:1 ratio, as described abovein Example 1) is placed in the vitreous of both of the woman's eyesusing a trocar. After about 2 days, the woman begins to notice a changein her eyes, presumably due to a decrease in intraocular pressure. Theloss of vision is prevented for about five months after the implantprocedure.

Example 4 Treatment of Ocular Conditions with Various Active Agents

An implant can be formulated with various active agents, including theagents described herein, following the procedures in the Examples above.These implants can provide an extended therapeutic treatment of anocular condition, that is a therapeutic effect during a period of timeduring release of the active agent or after release of all of the activeagent from the implant and during which there is no longer a therapeuticamount of the active agent present at the ocular site at which theimplant was placed. Thus, an implant can be prepared containing analpha-2 adrenergic receptor agonist, such as clonidine, apraclonidine,or brimonidine (available from Allergan, Irvine, Calif. as brimonidinetartrate ophthalmic solution, under the tradename Alphagan-P®). Thus,for example, a brimonidine extended therapeutic treatment implant can beimplanted into an ocular site (i.e. into the vitreous) of a patient withan ocular condition for a desired extended therapeutic effect. Theimplant may contain from about 50 μg to about 500 μg of Alphagan orAlphagan-P depending on the size of the implant. The brimonidineextended therapeutic treatment implant can be implanted into an ocularregion or site (i.e. into the vitreous) of a patient with an ocularcondition for a desired therapeutic effect. The ocular condition can bean inflammatory condition such as uveitis or the patient can beafflicted with one or more of the following afflictions: maculardegeneration (including non-exudative age related macular degenerationand exudative age related macular degeneration); choroidalneovascularization; acute macular neuroretinopathy; macular edema(including cystoid macular edema and diabetic macular edema); Behcet'sdisease, diabetic retinopathy (including proliferative diabeticretinopathy); retinal arterial occlusive disease; central retinal veinocclusion; uveitic retinal disease; retinal detachment; retinopathy; anepiretinal membrane disorder; branch retinal vein occlusion; anteriorischemic optic neuropathy; non-retinopathy diabetic retinal dysfunction,retinitis pigmentosa and glaucoma. The implant(s) can be inserted intothe vitreous using the procedure such as trocar implantation. Theimplant can release a therapeutic amount of the active agent to provideand retain a therapeutic effect for an extended period of time tothereby treat a symptom of an ocular condition. For example, the implantmay be effective to improve visual acuity, visual contract sensitivity,or both.

Example 5 Use of Intraocular Alpha-2 Adrenergic Receptor AgonistsImplants to Enhance, Restore and/or Improve Visual Acuity

Experiments were carried out with intraocular implants in mammalianeyes. Thus, degradable polymer implants containing as active agent anintraocular alpha-2 adrenergic receptor agonist were placed in thevitreous of both normal and damaged or diseased (model system) rabbiteyes. The results of the experiments showed that an intraocular alpha-2adrenergic receptor agonist implant can: (1) enhance visual acuity innormal eyes, and; (2) restore visual acuity in diseased or damaged eyes.

Formulation #17 of Example 1 was used in these Example 5 experiments.Thus, the implants used comprised the alpha-2 agonist brimonidinetartrate formulated as a solid, biodegradable rod (weighing about 1 mg)to form a sustained release drug delivery device (i.e. an implant). Theimplant consisted of 200 μg brimonidine tartrate and 800 μg of apoly-lactide co-polymer mixture of resomers R203 and R206 in a 1:1weight ratio. The placebo implant was a 1 mg rod implant made of apoly-lactide co-polymer mixture of resomers R203 and R206 in a 1:1weight ratio.

When the implant was administered intravitreally in rabbits we foundthat enhanced or normalized visual acuity resulted. Visual acuity wasmeasured as a sweep visual evoked potential threshold. Improved visualacuity was measured in each of three different ocular conditions:

1) in normal rabbit eyes;

2) in VEGF damaged rabbit eyes. These rabbit eyes were treatedintravitreally with 500 ng of VEGF to induce optic nerve head swelling,retinal vessel leak, dilation and tortuosity, to thereby simulatedisease aspects common to ocular conditions such as macular edema, opticnerve head edema, diabetic retinopathy, and neovascularization, and;

3) in rabbit eyes with outer retina injury induced by a transientischemic event with elevated IOP 8 months prior to use of the implant.

Significantly: (a) in ocular condition 2) above, the implant improvedvisual acuity without reducing the vasculopathy (tortuosity, leak,dilation) associated with the VEGF treatment, and; (b) in ocularcondition 3) above, the implant improved visual acuity without changingthe clinical appearance of the retina in these ischemic damage eyes, asassessed by color fundus photography. This indicates that the alpha 2agonist active agent released by the implant caused an augmentation ofthe tonic activity of the functioning retinal neuronal cells thatremained following the induced injury or disease state in ocularconditions 2) and 3). It can be hypothesized that the remaining normalcells functioned better to compensate for and to improve vision eventhough the disease state was still present.

Procedure for Inducing Retinal Ischemic Injury

Rabbits were anesthetized with isofluorane, and prepared for unilateralacute retinal ischemia by raising IOP in the OD eye by 120 mm Hg for 45minutes. To accomplish this, a reservoir with PBS was suspended 65inches above the eye and connected to a 30 gauge needle inserted throughthe cornea into the anterior chamber. A drop of topical anesthetic(proparacaine) was placed upon the cornea prior to needle insertion.And, a drop of anti-inflammatory agent (Pred-G) was placed onto thecornea immediately following needle removal.

Procedure for Measuring Visual Acuity

Visual acuity was measured in conscious rabbits using sweep visualevoked potential (swVEP). swVEP is an electrophysiological technique forassessing visual acuity typically used young children who can not readthe Snellen eye charts. Pattern reversal images of increasing spatialfrequency are projected onto the macula while simultaneously recordingelectrical activity (VEP) from the scalp. Images with lower spatialfrequency generate large signals which get smaller as the spatialfrequency increases, until signal=noise; this threshold is the visualacuity. The procedure in rabbits requires first implanting permanentelectrodes on the scalp to enhance signal strength and allow recordingfor the same position from follow-up visits. After two-weeks to allowfor healing, the visual acuity measurements can be made.

Rabbits were anesthetized with ketamine and xylazine for theimplantation procedure. The scalp was aseptically prepared and implantedwith four stainless steel screws (#0-80×⅜). Two active electrodes wereplaced at 6 mm on either side of the midline, 6 mm above bregma; 1ground electrode was placed at midline, 6 mm above the activeelectrodes; and, 1 reference electrode was placed at midline, 6 mm abovethe grounding electrode.

For the acuity test, eyes were fully dilated with 1% tropicamide and 10%phenylephrine. The rabbits were placed in stainless steel restrainersthat allowed projection of the pattern-reversal images onto the visualstreak. The rabbits were fully-conscious. Images were projected via aspecially-designed fundus camera stimulator under control of thePowerDiva software version 1.8.5. Each rabbit was placed so that its eyewas located at 50 mm in front of camera which is equivalent to 50 cmfrom a 21 in CRT monitor. Recording electrodes were connected to GrassNeurodata Acquisition System (Model 12CA) with the followingspecifications:

Channel 1 for OD eye and Channel 2 for OS eye.

Filter range between 3 to 100 Hz.

Amplification: 50K

Line frequency filter=OFF.

Vertical steady-state pattern-reversal sweep stimulus at spatialfrequency range of 0.1 to 5 cycles per degree at a temporal frequency of7.5 Hz were applied to the eye at mean luminance of 600 cd/m2 andcontrast of 80%. Five to 40 trials, 10 secs each, were collected fromeach eye. The number of trials was based on the signal-to-noise ratio.Trials were averaged and the threshold (visual acuity) was determined bysoftware or manual fitting at signal-to-noise ration no less than 2.5.Threshold values were then normalized by expressing as a percent of thecontralateral eye.

The effect on visual acuity of the brimonidine implant vs placebo wasstudied in three different conditions on six different group of rabbiteyes, as explained below. For each of the three groups of rabbits theimplants were placed in the vitreous using The implants are insertedinto the vitreous using the applicator (with a 22 gauge needle) setforth in U.S. patent application Ser. No. 11/021,947, filed Dec. 23,2004.

(A) In the first study two groups of rabbits with normal (untreated)eyes were used. One brimonidine implant was intravitreally implantedinto each left eye of each rabbit of group 1 (N=7). One placebo implantwas intravitreally implant into the left eye of each rabbit of group 2(N=7) using the same procedure. The right eye of each rabbit in bothgroups 1 and 2 was not treated and served as controls to normalize thedata obtained. Visual acuity was measured in both eyes of the rabbits inboth groups and is set forth in FIG. 15 as a percent of the visualacuity of the contralateral (right) normal eye.

FIG. 15 shows the effect of the brimonidine implant and of the placeboimplant on visual acuity in normal eyes of rabbits. The FIG. 15 resultswere recorded two weeks after implantation of either the brimonidineimplant or the placebo implant and show that the placebo implant did notcause a significant visual acuity change. Thus, the placebo implantcaused visual acuity to change only by 1.5%±6%. However, use of thebrimonidine implant clearly caused a significant enhancement of visualacuity in normal rabbit eyes. Thus, the brimonidine implant causedvisual acuity to improve by 44%±12% (up to a 56% vision improvement in anormal eye). A comparison of the responses to the placebo implant and tothe brimonidine implant with an unpaired Student's ‘T’ test showstatistical difference with a p value of 0.003.

(B) In the second study two groups of rabbits eyes were used. Onebrimonidine implant was intravitreally implanted into each left eye ofeach rabbit of group 1 (N=7). One placebo implant was intravitreallyimplant into the left eye of each rabbit of group 2 (N=7) using the samepars plana insertion procedure. The right eye of each rabbit in bothgroups 1 and 2 was not treated and served as controls to normalize thedata obtained. Two weeks after implantation each implanted eye wasintravitreally administered 500 ng of vascular endothelial factor (VEGF)(obtained from R&D Systems as product number 293-VE-50) as a 50 μlbolus. Visual acuity was measured in both eyes of the rabbits in bothgroups and is set forth in FIG. 16 as a percent of the visual acuity ofthe contralateral (right) eye.

FIG. 16 shows the effect of the brimonidine implant and of the placeboimplant on visual acuity in the VEGF treated rabbit eyes. The FIG. 16results were recorded three weeks after implantation of either thebrimonidine implant or the placebo implant and one week after VEGFadministration. The FIG. 16 results show that the placebo implant, VEGFtreated eyes had a visual acuity deficit of about 25%±4%. The FIG. 16results also show that the brimonidine implant, VEGF treated eyes had avisual acuity improvement of about 14%±8% (up to 22% vision improvementin an eye with a vasculopathy). Thus, use of the brimonidine implantnormalized visual acuity in eyes treated with VEGF despite the presenceof vasculopathy. The brimonidine implant did not reduce the vasculopathyinduced by VEGF, but did reduce the neurosensory retina deficit inducedby the VEGF treatment. A comparison of the responses to the placeboimplant and to the brimonidine implant with an unpaired Student's ‘T’test showed statistical difference with a p value of 0.0007.

(C) In the third study two groups of rabbits' eyes were used. Onebrimonidine implant was intravitreally implanted into each left eye ofeach rabbit of group 1 (N=5). One placebo implant was intravitreallyimplant into the left eye of each rabbit of group 2 (N=5) using the sameprocedure. The right eye of each rabbit in both groups 1 and 2 was nottreated and served as controls to normalize the data obtained. At dayzero ischemic injury was induced in the left eye of each rabbit in bothgroups. Thirty two weeks later each ischemic injury left eye of eachrabbit in group 1 was implanted with the placebo implant and each lefteye of each ischemic injury left eye of each rabbit in group 2 wasimplanted with the brimonidine implant. At week forty four visual acuitywas measured in both eyes of the rabbits in both groups and is set forthin FIG. 17 as a percent of the of the visual acuity of the contralateralnormal or untreated (right) eye.

FIG. 17 shows the effect of the brimonidine implant and of the placeboimplant on visual acuity in eyes of rabbits with existing injury from anischemic event. Histology shows that this procedure results in outerretina injury to photoreceptors, the RPE and associated tissues. Rabbitswith a visual acuity deficit in the left resulting from the transientischemic procedure were randomized into two groups. Data are expressedas a percent of the contralateral normal eye. The FIG. 17 results wererecorded twelve weeks after implantation with either the brimonidineimplant or the placebo implant and eleven months after the inducedischemic event to each implanted eye. The FIG. 17 results show that theplacebo implant, ischemic injury eyes has a visual acuity decrease of37%±8%. The FIG. 17 results also show that the brimonidine implant,ischemic injury eye had a visual acuity improvement of 14%±9%. Thus, useof the brimonidine implant restored or improved visual acuity in rabbiteyes with an outer retina (induced ischemic) injury. A comparison of theresponses to the placebo implant and the brimonidine implant with anunpaired Student's ‘T’ test showed a statistical difference with a pvalue of 0.001

These experiments showed that a locally (i.e. intravitreally)administered alpha-2 adrenergic receptor agonist can be used to improvevision (enhance visual acuity) in normal eyes. These experiments alsoshowed that a locally (i.e. intravitreally) administered alpha-2adrenergic receptor agonist can be used to improve, repair or restorevision in eyes with an ocular condition such as an inflammatory,neovascular, tumor, vascular occlusive, and/or optic nerve disease orcondition, including glaucoma.

A separate group of rabbits were studied to obtain pharmacokinetic data.In these rabbits the implants were inserted using a surgicalintravitreal implantation procedure performed as follows: a conjunctivalincision was made, and a sclerotomy was performed with a 20-gauge MVRblade. The sclerotomy was 3 mm from the limbus and lateral to the dorsalrectus muscle between the 10 and 12 o'clock positions on the right eye,and between the 12 and 2 o'clock positions on the left eye. Using asterile forceps, the test article was inserted through the sclerotomy.The sclerotomy was closed with 9-0 Prolene suture material. A sterileocular lubricant was applied to the eye following the implantationprocedure. Blood was collected from rabbits prior to euthanasia/necropsyon Days 8, 31, 58, 91, 136, or 184. The aqueous humor, vitreous humor,lens, retina and plasma samples from the rabbits were analyzed by usingliquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS)methods.

Table 3 shows the data obtained in this pharmacokinetic study.Intraocular brimonidine concentrations in the aqueous humor,iris-ciliary body, lens, retina, vitreous humor and plasmaconcentrations at various times (“Days” in the left hand side column ofTable 3) after intravitreal (into the midvitreous) implantation of the200 μg brimonidine implant (Formulation #17) in albino rabbit eyes wasmeasured. As shown by Table 3, after intravitreal implantation of theFormulation #17 brimonidine implant:

1. brimonidine was not detectable at any time point in the aqueous humorof the rabbit eyes implanted with the brimonidine implant;

2. the brimonidine had a posterior clearance as opposed to an anteriorclearance after release from the intravitreal implant, as shown by thehigher retinal concentration;

3. detectable levels of brimonidine were released from the implant intothe vitreous over at least a ninety day period;

4. therapeutic levels of the brimonidine existed in the retina for abouttwice as long at the implant released brimonidine from the implant:brimonidine was present in the retina for at least 84 days, although allthe brimonidine had been released from the implant after about 91 toabout 120 days;

5. the implant allowed an intra-retinal depot of brimonidine to beformed.

TABLE 3 Intraocular brimonidine concentrations Aqueous Iris-ciliary LensRetina Vitreous Plasma Day humor (ng/mL) body (ng/g) (ng/g) (ng/g) humor(ng/mL) (ng/mL) 8 NC 942 (3010)^(d) 45.1 ± 13.4 3630 ±

47.2 ± 13.1 0.092 (0, 0.184)    31 NC 25.9 ± 9.11 17.0 ± 3.92  35.3 ±15.5 9.35 ± 6.25^(b) 0.0575 (0, 0.115)    58 NC 69.4 ± 55.3 17.9 ±12.5^(b)  122 ± 57.3^(a)  5.6 ± 3.24^(b) 0.255 (0.208, 0.302) 91 NC 42.9± 18.7^(c) 50.1 ± 14.8  488 ± 471^(b) 59.3 ± 43.2 NC 136 NC  107 ± 41.516.2 ± 12.3^(a)  22.6 ± 5.9 NC NC 184 NC NC 1.18 ± 0.71^(b)  59.8 ±35.0^(b) NC NC NC, not calculable because >50% of concentrationscontributing to mean were BLQ (below limit of quantification). Dataexpressed as mean ± SEM (N = 4 eyes and N = 2 plasma per sampling time).BLQ = Below limit of quantitation (aqueous and vitreous humor: <10ng/mL; lens, retina and iris-ciliary body: <0.5 ng; plasma: <0.05ng/mL). ^(a)N = 4. One sample was BLQ (included in the mean calculationas zero). ^(b)N = 4. Two samples were BLQ (included in the meancalculation as zero). ^(c)N = 3. One sample was not determined (notincluded in the mean calculation). ^(d)N = 2. Two samples were ALQ(above limit of quantification) (estimated mean value in parentheses).

indicates data missing or illegible when filed

To conclude, our results show that an alpha-2 agonist (non-selective orreceptor subtype selective) intravitreal implant can be used to enhance,repair, restore or improve visual acuity in mammalian eyes.

Significantly, our experiments showed that an intravitreal brimonidineimplant can be used to improve visual acuity in both normal eyes and indiseased eyes. The results presented herein show that in VEGF treatedeyes show that the implant can be used as a prophylactic to prevent afuture vision loss. The results presented herein in damaged/diseased(i.e. ischemic) eyes show that an implant can be used to improve visualacuity in an eye without remission or disappearance of the eyedamage/disease. That is, the implant appears to cause the remainingnormal retinal cells to function better to compensate and improvevision, even though the eye damage/disease has not been reduced as toit's' physical extent in the retina.

Implants within the scope of our invention can be used:

1. as a prophylaxis to mitigate against impending retinal neurosensorydysfunction in a variety of ocular conditions, including retinaldisorders in patients that have a predisposition to or risk factorsassociated with a retinal disorder.

2. as a therapeutic (alone or in combination with one or more additionalactive agents) to treat posterior ocular conditions, such as retinaldiseases associated with degeneration of the retina, such as a maculadegeneration (such as an age related macular degeneration), an ocularedema, such as a macular edema, a vascular occlusive condition, an opticor retinal neuropathy, and/or a retinal tumor. For example, an implantcan be made comprising an alpha 2 agonist to lower IOP and/or to improvevisual acuity and a steroid (such as dexamethasone or triamcinolone) toreduce inflammation.

3. as a therapeutic (alone or in combination with one or more additionalactive agents) useful in retinal diseases and disorders with detachmentof the retina.

4. as a therapeutic (alone or in combination with one or more additionalactive agents) useful in surgical retinal procedures that requirevitrectomies and manipulation that can have a negative impact of theretina.

5. as a therapeutic (alone or in combination with one or more additionalactive agents) to treat retinal diseases that have a nutritionaldeficiency, such as a vitamin A deficiency

6. as a therapeutic (alone or in combination with one or more additionalactive agents) to treat retinal injury from accidental light exposure,such an operating microscope light or industrial lasers.

7. as an adjunct with steroids for treating retinal diseases, wheresteroids are used to reduce ocular inflammation and macular or opticnerve edema.

8. as an adjunct to photodynamic therapy (PDT) where PDT is used totreat retinal conditions associated with leakage from retinal andrelated tissue vessels.

9. as an adjunct to other types of electromagnetic radiation such aslaser photocoagulation used to treat macula edema or neovascularization,and transpupillary thermal therapy (TTT) that is used to treat choroidalneovascularization (CNV).

10. as an adjunct to radiation therapy or chemical therapy that causesmaculopathy and papillopathy when used to treat ocular tumors such asmacular retinoblastoma and choroidal osteoma.

11. as an adjunct to electromagnetic radiation and steroids used totreat edema and neovascular abnormalities of the eye.

Example 6 Use of Two Different Intravitreal Brimonidine Implants toTreat Acute Rhegmatogenous Macular-Off Retinal Detachment

Patients are implanted with either a 50 μg and 200 μg brimonidineposterior segment (i.e. intravitreal) implant to treat acuterhegmatogenous macular-off retinal detachment. The patients experienceat least a 15-letter increase from baseline in the study eye using theEarly Treatment Diabetic Retinopathy Study (ETDRS) method). The 200 μgbrimonidine posterior segment implant is more effective than the 50 μgbrimonidine posterior segment implant in achieving an improvement inbest-corrected visual acuity (BCVA) (as measured by the proportion ofpatients experiencing at least a 15-letter increase from baseline in thestudy eye using the Early Treatment Diabetic Retinopathy Study (ETDRS)method). The 50 μg and 200 μg brimonidine posterior segment implantshave acceptable safety profiles

The patients are seen for a baseline and randomization visit on day (O),and at months 1, 3, 6, 9, 12 (masked phase) and 15, 18, and 24 months(extension phase). Additional visits on day 1 and day 7 following anyre-treatments are designated as safety visits. Fifty five patients areenrolled. At least one eye of each patient has acute rhegmatogenousmacular-off retinal detachment eligible for repair by scleral buckle andlaser photocoagulation

The Inclusion criteria for the patients include: eighteen years of ageor older; best corrected E-ETDRS visual acuity score of >=20 letters(i.e., approximately 20/400 or better) and <=65 letters (i.e.,approximately 20/50 and worse); diagnosis of acute rhegmatogenousmacular-off retinal detachment in one eye. The detachment has occurredwithin twelve hours of presentation, and in the opinion of theinvestigator, can be repaired by scleral buckle placement and externallaser photocoagulation of the retinal break without the anticipated needfor vitrectomy or pneumatic retinopexy. The surgical repair is plannedwithin 48 hrs of the detachment; media clarity; pupillary dilation, and;patient cooperation sufficient for adequate fundus photographs.

The study formulations are: (1) Formulation #17 of Example 1 (a solid,biodegradable rod (weighing about 1 mg implant consisted of 200 μgbrimonidine tartrate and 800 μg of a poly-lactide co-polymer mixture ofresomers R203 and R206 in a 1:1 weight ratio, and; (2) a solid,biodegradable rod (weighing about 1 mg implant consisted of 50 μgbrimonidine tartrate and 950 μg of a poly-lactide co-polymer mixture ofresomers R203 and R206 in a 1:1 weight ratio.

A significant number of the patients show an increase of 15 letters ormore from baseline of BCVA using the ETDRS method at 6 months in thestudy eye. Hence, an intravitreal brimonidine implant can be used totreat acute rhegmatogenous macular-off retinal detachment.

Example 7 Use of Two Different Intravitreal Brimonidine Implants toTreat Chronic Retinal Injury

Patients are implanted with either a 50 μg and 200 μg brimonidineposterior segment (i.e. intravitreal) implant to treat chronic retinalinjury. The patients experience an improvement in best-corrected visualacuity (BCVA) (as measured by the proportion of patients experiencing atleast a 15-letter increase from baseline in the study eye using theEarly Treatment Diabetic Retinopathy Study (ETDRS) method). The 200 μgbrimonidine posterior segment implant is more effective than the 50 μgbrimonidine posterior segment implant in achieving an improvement inbest-corrected visual acuity (BCVA) (as measured by the proportion ofpatients experiencing at least a 15-letter increase from baseline in thestudy eye using the Early Treatment Diabetic Retinopathy Study (ETDRS)method). The 50 μg and 200 μg brimonidine posterior segment implantshave acceptable safety profiles.

The patients are seen for a baseline and randomization visit on day (0),and at months 1, 3, 6, 9, 12 (masked phase) and 15, 18, and 24 months(open label). Additional visits on day 1 and day 7 following anyre-treatments are designated as safety visits. At least one eye of eachpatient has retinal injury at baseline of one or more of the followingtypes: tapetoretinal degeneration, macular ischemia due to diabeticmaculopathy or prior rhegmatogenous macular-off retinal detachment.

The inclusion criteria for the patients include: a least eighteen yearsof age; diagnosis of chronic retinal injury related to tapetoretinaldegeneration, macular ischemia due to diabetic maculopathy ormacular-off retinal detachment in at least one eye (the study eye); forpatients with tapetoretinal degeneration the diagnosis is based on bothclinical, visual field and electroretinographic findings and the visualfield loss is within the central 10 degrees; for patients with macularischemia due to diabetic maculopathy the best corrected E-ETDRS visualacuity score is >=35 letters (i.e., approximately 20/200 or better) and<=65 letters (i.e., approximately 20/50 or worse), and the decreasedvisual is directly related to the ischemia and not due to macular edemaor previous laser photocoagulation; for patients with macular-offdetachment: the macula detachment has not been present longer than 48hours prior to repair, the repair of the detachment has occurred atleast 6 months before the baseline visit, the best corrected E-ETDRSvisual acuity score is >=35 letters (i.e., approximately 20/200 orbetter and <=65 letters (i.e., worse than approximately 20/50), thevisual acuity is stable (is within one line of Snellen acuity) for atleast 3 months, the repair of the detachment has been deemed an anatomicsuccess; media clarity; pupillary dilation, and; patient cooperationsufficient for adequate fundus photographs.

The study formulations are: (1) Formulation #17 of Example 1 (a solid,biodegradable rod (weighing about 1 mg implant consisted of 200 μgbrimonidine tartrate and 800 μg of a poly-lactide co-polymer mixture ofresomers R203 and R206 in a 1:1 weight ratio, and; (2) a solid,biodegradable rod (weighing about 1 mg implant consisted of 50 μgbrimonidine tartrate and 950 μg of a poly-lactide co-polymer mixture ofresomers R203 and R206 in a 1:1 weight ratio.

A significant number of the patients show an increase of 15 letters ormore from baseline of BCVA using the ETDRS method at 12 months in thestudy eye. Hence, an intravitreal brimonidine implant can be used totreat chronic retinal injury.

Example 8 Manufacture and In Vitro Testing of Drug Delivery SystemsContaining an Alpha-2 Adrenergic Receptor Agonist and a BiodegradablePolymeric Matrix Comprising Ester End and Acid End Polymers

Introduction

It is known to prepare biodegradable polymer implants capable ofreleasing an active agent. See for example Lewis, D., Controlled Releaseof Bioactive Agents from Lactide/Glycolide Polymers in Drugs andPharmaceutical Sciences, Vol. 45, “Biodegradable Polymers as DrugDelivery Systems”, edited by Chasin M., et al., pages 1-35 (1990), and;de Jong S., et al., New insights into the hydrolytic degradation of poly(lactic acid): participation of the alcohol terminus, Polymer 42 (2001);2795-2802.

An implant (synonymously a drug delivery system) can have undesirablerelease characteristics. Indeed, one of the most difficult andproblematic aspects of implant technology is the discovery anddevelopment of an implant with desirable and consistent active agentrelease characteristics. Implant release characteristics (the releaseprofile) depends upon a multitude of factors including the chosen activeagent (including it's solubility, reactivity and labiality), theselection of a particular polymer or polymers from the near infinitenumber of different polymers and polymer combinations available, and themanufacturing process by which the implant is made.

Undesirable implant release characteristics can include an initial burstrelease of the active agent and/or a lag time in the release of theactive agent, both illustrated by FIG. 18. Such undesirable implantrelease characteristics can cause overdosing or underdosing of thepatient with the active agent with resulting reduced therapeuticefficacy of the implant. Although an implant with a burst release orwith a lag time may have utility in some circumstances, generally adesirable implant release characteristic is a linear rate of release ofthe active agent, thereby providing a constant or relatively constantdosing of the therapeutic agent to the patient.

Thus, an implant intended for intraocular administration which comprisesan alpha-2 adrenergic receptor agonist (as the active agent) and abiodegradable polymeric matrix can exhibit a burst effect or asignificant lag time after ocular implantation or insertion of theimplant before release of a therapeutically effective amount of thealpha-2 adrenergic receptor agonist from the polymeric matrix of theimplant (for example into the vitreous) takes place. A burst can be dueto having too much active drug incorporated at or near the surface of animplant, and a lag time to having too little of the active agentincorporated at or near the surface of the implant. For example, for abiodegradable implant, having more than about 25% of the active agentwithin the polymeric matrix which comprises up to about the top 15% asmeasured from the exterior to the center of the implant) of the volumeof the implant can result in a burst effect. Concomitantly, for abiodegradable implant, having less than about 15% of the active agentwithin the polymeric matrix which comprises up to about the top 25% (asmeasured from the exterior to the center of the implant) of the volumeof the implant can result in a lag time.

Additionally, a burst effect or a lag time before release of atherapeutically effective amount of the alpha-2 adrenergic receptoragonist from the polymeric matrix of the implant (for example into thevitreous) can be due to a selection of polymer or polymers forconstituting the polymeric matrix of the implant which polymer orpolymers have characteristics which do not permit the active agent to bereleased with a linear or substantially linear release profile. Althoughbroad or general polymer characteristics are known (see eg example Lewis(1990) and de Jong S. (2001) supra) there is a near infinite variety ofdifferent polymer or polymer combinations (each with their owndegradation rates, pore forming characteristics, reactivities,degradation pathways, intermediates, by products, active agent-polymerassociation characteristics, etc) which can be used to constitute thepolymeric matrix of the implant.

The size and weight of implants intended for intraocular administrationis significantly limited by the dimensions of intraocular spaces andpotential intraocular spaces. Additionally, even when it may bephysically possible to insert, implant or inject an implant to or into aparticular intraocular site, considerations such as a desire to reduceinjury to sensitive ocular tissues at the site of and adjacent to thesite of administration site, and to not interfere with vision canrequire the implant to be less than it's maximum pp size and weight.Ocular tissue injury can result in inflammation, pain, increased healingtime and reduced visual acuity. In light of these considerations, animplant intended for example for intravitreal administration preferablyhas no dimension greater than about 20 mm and weighs less than about 5mg. For our purposes, the weight of the implant is the weight of theactive agent incorporated into the implant plus the weight of thepolymers or polymers which comprise the polymeric matrix of the implant.Having too little of the active agent incorporated into the implant canresult in having too little implant surface active agent (that is havingonly a minute absolute weight amount of the active agent present withina few microns of the exterior surface of the implant). Implant surfaceactive agent is available for immediate or rapid (i.e. within the firstday or two after implantation) release from the polymeric matrix.Additionally, having a low amount of implant surface soluble activeagent can delay implant surface pore formation which thereby slows therelease of the active agent from deeper within the implant. To addressthis problem pore forming additives have been used to improve theinitial release of an active agent from a biodegradable implant, butunfortunately use of pore forming additives can result in a significantdecrease in the duration of the time over which a therapeuticallyeffective amount of the active agent is released from the implant.

Summary

With an awareness of these problems and deficiencies of prior implantsand in light of the considerations above, we have developed implantsintended for the treatment of ocular conditions comprising an alpha-2adrenergic receptor agonist (as the active agent) and a biodegradablepolymeric matrix with the following desirable characteristics:

1. No burst effect and as well no or a nominal lag time after ocularimplantation or insertion of the implant before release of atherapeutically effective amount of the alpha-2 adrenergic receptoragonist from the implant occurs. Generally, we found that a suitableinitial release of the active agent from the implant as well as along-term sustained release of the active agent from the implant couldbe achieved by particular novel selections of hydrophobic poly(D,L,-lactide) polymers and/or hydrophilic poly(D,L,-lactide-co-glycolide) polymers to comprise the polymeric matrix ofthe implant.

2. high dose implants, that is implants which comprise more than 4weight percent (wt %) of a biologically active alpha-2 adrenergicreceptor agonist. The remaining 96 wt % or less of the implant willtypically comprise one or more biocompatible and biodegradable polymers.

3. absence of pore forming additives or other release rate modulators ormodifiers.

4. sustained release of a therapeutic amount of an alpha-2 adrenergicreceptor agonist from the biodegradable polymeric matrix over a periodof at least 115 days (about 4 months).

5. substantially linear (i.e. first order release rate kinetics) releaseof an alpha-2 adrenergic receptor agonist from the biodegradablepolymeric matrix of the implant over a period of time of from about 20days to about 50 days. “Substantially linear release” means that themeasured amount of the rate of alpha-2 adrenergic receptor agonistrelease from the biodegradable polymeric matrix of the implant does notvary by more than 50% over a three day period, preferably does not varyby more than 30% over a seven day period and most preferably does notvary by more than 20% over a ten day period.

Thus, we made brimonidine tartrate containing sustained release polymerimplants with exhibited desirable in vitro release profile of thebrimonidine tartrate¹. Generally the release profiles showed no or areduced initial drug burst and as well sustained release profile of theactive agent over a period of at least about 115 days. Our novel implantformulations contained hydrophobic ester end-capped poly (D,L-lactide)homopolymers, ester end-capped poly (lactide-co-glycolide) copolymers,and a minor amount of acid end-capped poly (D,L,-lactide-co-glycolide)polymer. An uncapped polymer (such as a PLGA polymer) has a freecarboxyl group at the polymer terminus. Without wishing to be bound bytheory we believe that these desirable implant release characteristicswere obtained because the polylactide component of the polymers used canprovide an implant polymeric matrix with a relatively long hydrolysishalf-life as well as a durable polymeric matrix capable of retaining theactive agent within the implant polymer matrix for an extended period oftime. Additionally, the acid-end capped lactide-glycolide polymer canact to speed initial active agent drug release from the implant byenhancing early water penetration into the implant by raising thesurface energy of the implant. The acid-end capped lactide-glycolidepolymer can also create a more porous implant as it swells and is erodedfrom the implant because its hydrolysis rate is much faster than that ofthe polylactide. ¹Brimonidine tartrate is a relatively selective alpha-2adrenergic agonist approved for ophthalmic use. The chemical name ofbrimonidine tartrate is5-bromo-6-(2-imidazolidinylideneamino)quinoxaline L-tartrate. It is anoff-white, pale yellow to pale pink powder. In solution, brimonidinetartrate has a clear, greenish-yellow color. Brimonidine tartrate has amolecular weight of 442.24 as the tartrate salt and is water soluble (34mg/ml). The molecular formula is C₁₁H₁₀BrN₅.C₄H₆O₆.

Experiments

We carried out experiments to make and to test our particular drugdelivery systems comprising an alpha-2 adrenergic receptor agonist and abiodegradable polymeric matrix comprising both ester end capped and acidend capped polymers. In particular we made and tested in vitrobrimonidine tartrate containing sustained release polymer implantsintended for intravitreal administration to treat an ocular condition.

Examples of three implants formulation we made comprising an alpha-2adrenergic receptor agonist and a biodegradable polymeric matrix areshown in Table 4. The in vitro release rates of the three Table 4implants over a 21 day period (in phosphate buffered saline (“PBS”) atpH 7.4 and 37° C. is shown by FIG. 19.

As shown in Table 4 and in FIG. 19, a particularly advantageous implantformulation made was formulation number 7746-073. Implant formulation7746-073 comprised 12 wt % brimonidine tartrate, 53 wt % R2035 (an esterend-capped poly (D,L,-lactide) polymer), 25 wt % R208 (also an esterend-capped poly (D,L,-lactide) polymer), and 10% RG502H (an acidend-capped poly(D,L-lactide-co-glycolide) polymer). Resomers RG502H,R208 and R2035 can be obtained from Boehringer Ingelheim. RG502Hcomprises 48-52 mol % D,L-lactide and 48-52 mol % glycolide (asdetermined by NMR spectroscopy) and has an inherent viscosity at 25° C.of 0.16 to 0.24 dl/g. R208 has an inherent viscosity at 25° C. of 1.8 to2.2 dl/g. R2035 has an inherent viscosity at 25° C. of 0.25 to 0.35dl/g.

TABLE 4 Formulations of Three Brimonidine Tartrate Implants w/w, %Brimonidine Resomer Resomer Resomer Formulation No Tartrate R203S R208RG502H 7746-073 12 53 25 10 7702-020 15 60 25 0 7702-022 18 65 17 0

As shown by FIG. 19, in distinction to the implant formulations whichcontained only ester end-capped biodegradable polymers (formulations7702-020 and 7702-022), the implant formulation which contained amixture of ester end-capped and acid end-capped polymers (formulation7746-073) showed an essentially linear rate of release of thebrimonidine tartrate active agent in vitro over the two week period ofdays 7-21.

Additionally, as shown by FIG. 20, when the release of the active agentfrom formulation 7746-073 was observed over a longer period of time (inPBS at pH 7.4 and 37° C.), linear rates of release of the brimonidinetartrate active agent in vitro were observed during the time periodsfrom about day 30 to about day 45 (15 day period), from about day 50 toabout day 90 (40 day period) and from about day 100 to about day 115 (15day period).

We then made seven additional implant formulations each comprising analpha-2 adrenergic receptor agonist and various ratios of esterend-capped and acid-end capped biodegradable polymers as shown in Table5. In Table 5 R2035 and R208 are ester end-capped poly (D,L,-lactide)polymer resomer supplied by Boehringer-Ingelheim (Resomer), RG502H is anacid end-capped poly (D,L-lactide-co-glycolide) polymer and RG752 isanother ester end capped poly (D,L,-lactide-co-glycolide) polymer. APO40is an ester end-capped poly (D,L,-lactide) polymer with an inherentviscosity at 30° C. of 0.34-0.40 dl/g, available from Durect Corporation(Lactel). All inherent viscosities for all resomers set forth hereinwere measured using CHCl₃ as solvent.

The in vitro release rates of the seven Table 5 implant formulationsover a 14-26 day period (in PBS at pH 7.4 and 37° C.) is shown by FIG.21. Significantly, FIG. 20 shows linear release profiles of the Table 5Formulations, all of which contained acid end-capped polymer incombination with poly (D,L,-lactide) and poly (lactide-co-glycolide)polymers.

Thus, FIG. 21 shows that a linear release profile of the active agentcan be obtained with different combinations of ester end-capped and acidend-capped polymers, even when: (1) the active agent loading in thepolymeric matrix is varied between about 9 wt % and about 12 wt %, and;(2) the wt % of acid end-capped polymer compositions is varied fromabout 15 wt % to about 35 wt %, the remainder of the implant being acombination of two or three different ester end-capped polymers.

TABLE 5 Linear Release Brimonidine Tartrate Containing Formulations w/w,% Brimonidine Resomer Resomer Resomer Resomer Formulation No TartrateAPO40 R203S R208 RG502H RG752 7746-061A 12 25 0 25 15 23 7702-068A 10.931.8 0 22.7 16.4 18.2 7702-065A 12 35 0 35 18 0 7702-070A 12 25 0 25 1820 7702-058A 12 23 0 40 25 0 7702-62A  12 20 0 20 25 23 7702-054A 9 0 650 35 0

The polymeric implants in this study were made by melt extrusion in aDaca instruments microcompounder/extruder, but they can also be made bydirect compression. The implants made were rod-shaped, but they can bemade into any geometric shape by changing the extrusion or compressiondie.

Polymers were used as received from Boehringer Ingelheim (Resomer) orDurect (APO40) and the brimonidine tartrate was used as the salt. Thepolymers and brimonidine tartrate were combined in a Retsch ball-millcapsule with a ¼″ stainless steel ball; then the capsule was placed inthe Retsch mill (Type MM200) for 5 min at 20-cycles/min. The capsule wasremoved from the mill and the powder blend was stirred with a spatula.The capsule with the powder blend was mixed for 5 minutes on a Turbulamixer. The powder blend is inspected for homogeneity and the mixingprocedure is repeated if necessary.

The Daca microcompounder/extruder was setup according to themanufacture's instructions. The output of the Daca is fitted with alaser micrometer and a custom built puller to control the diameter ofthe extruded rod. The Daca was allowed to equilibrate to the extrusiontemperature; then the powder blend was manually fed into the extrusionscrews at a rate that maintained a constant load and torque. All theExample 8 and Example 9 implant filaments were made at extrusiontemperatures between 95° C. and 115° C. The filaments made were cut intoone-milligram rods approximately 6 mm long and each with a diameter ofabout 0.018 (about 0.46 mm). Extruded filament active agent release wasmonitored by HPLC in phosphate buffered saline pH 7.4 at 37° C.

Example 9 Manufacture and In Vitro Testing of Drug Delivery SystemsContaining Two Forms of an Alpha-2 Adrenergic Receptor Agonist and anEster End-Capped Biodegradable Polymeric Matrix

In Example 1 above filament shaped implants containing brimonidine freebase and brimonidine tartrate were made using the single polymerresomers R203, R206 or RG752. See Formulations 11-15 and 19-20 inTable 1. These Example 1 implants were made using a process with mixing,melting, pelletizing and melt extrusion steps.

In this Example 9 experiment we made implants containing brimonidinefree base and brimonidine tartrate using a combination of two differentpolymer resomers and a different and improved manufacturing process, ascompared to Example 1. The Example 9 sustained release implants havingimproved release profiles were made using the Example 8 process, that isby combining a brimonidine free base and a brimonidine tartrate followedby stirring, mixing, and melt extrusion of implant filaments. Thefilaments made were cut into one-milligram rods, approximately 6 mm longand each with a diameter of about 0.018 (about 0.46 mm).

In this Example 9, the polymer matrix consisted of two poly(D,L-lactide)PLA polymers, although many different types of polymers can be used. Theimprovement in the release kinetics is primarily a function of thephysicochemical properties of the brimonidine free base and thebrimonidine tartrate. Therefore the substantially linear active agentrelease kinetics can be obtained with a variety of other polymers,beyond the two poly(D,L-lactide) PLA polymers used in this experiment.Because brimonidine tartrate is more water soluble than is brimonidinefree base, implants made with the tartrate can show a burst release dueto the availability of implant surface brimonidine tartrate. On theother hand, brimonidine free base is not water soluble, thereby makingthe implant more hydrophobic and delaying initial water permeation intothe implant and consequently therefore also the release of brimonidine,resulting in an observed lag before a therapeutic amount of thebrimonidine free base is released from the implant.

Based on the known solubility difference between brimonidine free baseand a brimonidine tartrate, the expected release rate of brimonidinefrom an implant which comprises both brimonidine free base and abrimonidine tartrate would be proportional to the amount of brimonidinetartrate in the formulation. Surprisingly, we discovered that an implantwhich comprises a combination of a brimonidine free base and abrimonidine tartrate has a synergistic release profile, that is therelease of brimonidine from the combination implant is faster than therelease rate of brimonidine from an implant which comprises only abrimonidine tartrate, with no brimonidine free base. FIG. 22 (in vitrorelease in PBS at pH 7.4 and 37° C.) shows that an implant whichcomprises a combination of the two forms (brimonidine free base andbrimonidine tartrate) shows a faster release than for either individualform of brimonidine in an implant by itself.

An embodiment of our new formulation can comprise brimonidine free base,brimonidine tartrate, and a biodegradable polymer, such as a hydrophobicbiodegradable polymer. The hydrophobic biodegradable polymer can be anester end-capped polymer. The hydrophobic biodegradable polymer can bean ester end-capped polymer, such as a hydrophobic, ester end-cappedpoly (D, L-lactide) homopolymer. Another embodiment of our invention cancomprise two hydrophobic, ester end-capped poly (D, L-lactide)homopolymers, as shown by the six Table 6 formulations. The polylactidescan have a relatively long hydrolysis half life and can provide a moredurable matrix to retain the active agent within the implant for anextended period.

As shown by FIG. 22 our implant formulation 7746-146 which comprised17.5% brimonidine free base, 17.5% brimonidine tartrate, 40% of theester end-capped poly (D,L,-lactide) polymer R203S, and 25% of the esterend-capped poly (D,L,-lactide) polymer R208 had a release profile whichwas faster than the formulations which contained only a brimonidine freebase or only a brimonidine tartrate (formulations 7746-118, 7746-141,7746-092A and 7746-142 in Table 5). Significantly, the 7746-146formulation had a first 20 day linear release profile followed by asecond, different 40 day linear release profile. Notably, formulation7746-147 which also comprised a combination of brimonidine free base andbrimonidine tartrate, showed an analogous active agent release profilebut with a different polymer ratio. All the formulations are summarizedin Table 6.

TABLE 6 Brimonidine Tartrate Containing Formulations w/w, % BrimonidineBrimonidine Resomer Resomer Formulation No Free Base Tartrate R203S R2087746-146 17.5 17.5 40 25 7746-118 0 35 40 25 7746-141 35 0 40 257746-147 17.5 17.5 55 10  7746-092A 0 35 55 10 7746-142 35 0 55 10

Example 10 Treatment of Elevated Intraocular Pressure with anIntravitreal Biodegradable Polymeric, Brimonidine Containing Implant

In Example 2 above it was disclosed that surgical placement into thevitreous of normotensive monkey eyes of a brimonidine containingbiodegradable polymeric implant did not lower IOP in the monkeys ascompared to placebo. See e.g. FIG. 12.

In this Example 10 we surprisingly show that surgical placement into thevitreous of hypertensive monkey eyes of a brimonidine containingbiodegradable polymeric implant does lower IOP in the monkeys ashypertensive compared to placebo hypertensive monkey eyes. See eg FIG.23. Example 10 can therefore be viewed as a further rendition of,supplement or extension of Example 3.

In this experiment we observed the effect on intraocular pressure (IOP)of brimonidine containing drug delivery system inserted into thevitreous of rabbit eyes (Dutch Belted rabbits). Elevated IOP was inducedin the rabbits by intracameral injection of Carbopol 934P, an ocularhypertensive (OHT) drug. IOP was measured using a hand heldpneumatonmeter.

Just as in Example 2, the extruded rod shaped Formulation 17 implant ofExample 1 was used as the brimonidine containing implant in this Example10 experiment. Thus, the active agent implant used weighed about 1 mgand comprised 20 wt % brimonidine tartrate (200 μg), 40 wt % resomerR203, and 40 wt % resomer R206 (800 μg polylactide polymer). TheFormulation 17 placebo 1 mg implant used comprised 50 wt % resomer R203and 50 wt % resomer R206 (1 mg polylactide polymer).

An implant was placed in the right eye only of twelve Dutch Beltedrabbits as follows. The rabbits were sedated by intravenous syringeinjection of Ketamine given in a dose of 15 mg per kg of rabbit weight(mg/kg) combined with acepromazine maleate 1 mg/kg. Next 1-2 drops ofocular Betadine was applied to the right eye as a topical disinfectantfollowed by 1-2 drops of Proparacaine applied to the right eye as atopical anesthetic.

The brimonidine containing implant was placed in the vitreous of sixright eyes and the placebo implant was placed in the vitreous of theother six eyes. The implants were placed by making an incision 4 mm fromthe limbus into the vitreous. In six right eyes an extruded rodbrimonidine implant was so placed. After implantation topical antibioticointment was applied to each right eye.

On the day after implantation, ocular hypertension was induced asfollows. The rabbits were sedated with sedated by intravenous syringeinjection of Ketamine 15 mg/kg combined with acepromazine maleate 1mg/kg. Next 1-2 drops of ocular Betadine was applied to the right eye asa topical disinfectant followed by 1-2 drops of Proparacaine applied tothe right eye as a topical anesthetic. Next 50 μl of 0.3% Carbopol 934Psolution at pH 4 was administered to each right eye by intracameralinjection, followed by application of topical antibiotic ointment to theright eye.

As shown by FIG. 23, as compared to placebo the brimonidine containingimplant lowered IOP in the hypertensive right eyes of the subjectrabbits for up to six weeks after DDS implantation.

All references, articles, publications and patents and patentapplications cited herein are incorporated by reference in theirentireties.

While this invention has been described with respect to various specificexamples and embodiments, it is to be understood that the invention isnot limited thereto and that it can be variously practiced within thescope of the following claims.

We claim:
 1. A biodegradable intraocular implant comprising brimonidinefree base in an amount in the range of 1 wt % to 50 wt %, and abiodegradable polymer, wherein the biodegradable polymer comprises anester end-capped biodegradable polymer and an acid end-cappedbiodegradable polymer.
 2. The implant of claim 1, wherein the implantcomprises from about 10% to about 91% ester end-capped biodegradablepolymer, from about 5 wt % to about 40 wt % acid end-cappedbiodegradable polymer, and from about 4 wt % to about 50 wt %brimonidine free base.
 3. The implant of claim 2, wherein the implantcomprises from about 25% to about 80% ester end-capped biodegradablepolymer, from about 10 wt % to about 40 wt % acid end-cappedbiodegradable polymer, and about 10 wt % to about 35 wt % brimonidinefree base.
 4. The implant of claim 2, wherein the implant comprisesabout 88 wt % ester end-capped biodegradable polymer, about 10 wt % acidend-capped biodegradable polymer, and about 12 wt % brimonidine freebase.
 5. The implant of claim 2, wherein the implant comprises fromabout 53 wt % to about 73% ester end-capped biodegradable polymer, fromabout 15 wt % to about 35 wt % acid end-capped biodegradable polymer,and from about 9 wt % to about 12 wt % brimonidine free base.
 6. Theimplant of claim 2, wherein the biodegradable polymer comprises morethan one ester end-capped biodegradable polymer.
 7. The implant of claim2, wherein the biodegradable polymer comprises more than one acidend-capped biodegradable polymer.
 8. The implant of claim 2, wherein theimplant has no or a nominal lag time after ocular implantation orinsertion of the implant before release of a therapeutically effectiveamount of the brimonidine free base from the implant occurs.
 9. Theimplant of claim 1 which does not include any pore forming additives,release rate modulators or release rate modifiers.
 10. The implant ofclaim 1, wherein the implant can exhibit a sustained release of thebrimonidine free base from the biodegradable polymeric matrix over aperiod of at least 115 days.
 11. The implant of claim 1, wherein theimplant exhibits a substantially linear release of the brimonidine freebase from the biodegradable polymeric matrix of the implant over aperiod of time of from about 20 days to about 50 days.
 12. A process formaking a biodegradable intraocular implant comprising: (a) mixing abrimonidine free base and a biodegradable polymer, wherein thebiodegradable polymer comprises an ester end-capped biodegradablepolymer and an acid end-capped biodegradable polymer; (b) heating themixture, and; (c) extruding the heated mixture, to thereby make abiodegradable intraocular implant.
 13. A biodegradable intraocularimplant made by the process of claim 12, wherein the brimonidine freebase is homogenously distributed throughout the implant.
 14. A methodfor treating an ocular condition selected from the group consisting of:macular degeneration, age related macular degeneration, non-exudativeage related macular degeneration, exudative age related maculardegeneration, choroidal neovascularization, retinopathy, diabeticretinopathy, acute and chronic macular neuroretinopathy, central serouschorioretinopathy, macular edema, cystoid macular edema, and diabeticmacular edema, acute multifocal placoid pigment epitheliopathy, Behcet'sdisease, birdshot retinochoroidopathy, syphilis, lyme disease,tuberculosis, toxoplasmosis, uveitis, intermediate uveitis, parsplanitis, and anterior uveitis, multifocal choroiditis, multipleevanescent white dot syndrome, ocular sarcoidosis, posterior scleritis,serpignous choroiditis, subretinal fibrosis, uveitis syndrome,Vogt-Koyanagi-Harada syndrome, retinal arterial occlusive disease,central retinal vein occlusion, disseminated intravascular coagulopathy,branch retinal vein occlusion, hypertensive fundus changes, ocularischemic syndrome, retinal arterial microaneurysms, Coat's disease,parafoveal telangiectasis, hemi-retinal vein occlusion,papillophlebitis, central retinal artery occlusion, branch retinalartery occlusion, carotid artery disease, frosted branch angitis, sicklecell retinopathy and other hemoglobinopathies, angioid streaks, familialexudative vitreoretinopathy, Eales disease, sympathetic ophthalmia,uveitic retinal disease, retinal detachment, eye trauma, laser inducedeye damage, photocoagulation, eye hypoperfusion during surgery,radiation retinopathy, bone marrow transplant retinopathy, proliferativevitreal retinopathy, appearance of epiretinal membranes, proliferativediabetic retinopathy, ocular histoplasmosis, ocular toxocariasis,presumed ocular histoplasmosis syndrome, endophthalmitis, toxoplasmosis,retinal diseases associated with HIV infection, choroidal diseaseassociated with HIV infection, uveitic disease associated with HIVInfection, viral retinitis, acute retinal necrosis, progressive outerretinal necrosis, fungal retinal diseases, ocular syphilis, oculartuberculosis, diffuse unilateral subacute neuroretinitis, myiasis,retinitis pigmentosa, systemic disorders with associated retinaldystrophies, congenital stationary night blindness, cone dystrophies,Stargardt's disease and fundus flavimaculatus, Bests disease, patterndystrophy of the retinal pigmented epithelium, X-linked retinoschisis,Sorsby's fundus dystrophy, benign concentric maculopathy, Bietti'scrystalline dystrophy, pseudoxanthoma elasticum, retinal detachment,macular hole, giant retinal tear, retinal disease associated withtumors, congenital hypertrophy of the RPE, posterior uveal melanoma,choroidal hemangioma, choroidal osteoma, choroidal metastasis, combinedhamartoma of the retina and retinal pigmented epithelium,retinoblastoma, vasoproliferative tumors of the ocular fundus, retinalastrocytoma, intraocular lymphoid tumors, punctate inner choroidopathy,acute posterior multifocal placoid pigment epitheliopathy, myopicretinal degeneration, acute retinal pigment epithelitis, the methodcomprising the step of intraocular administration of a biodegradableintraocular implant comprising brimonidine free base and a biodegradablepolymer, wherein the biodegradable polymer comprises an ester end-cappedbiodegradable polymer and an acid end-capped biodegradable polymer.